KingFisher systems can handle a broad range of applications across nucleic acids, proteins and peptides, cells and exosomes.

 

Different sample types and applications require different chemistries to extract the analyte. For example, blood, saliva, and buccal samples are concentrated sources of long DNA fragments, while cell-free DNA samples are high-volume, low-concentration sources of short fragments. It takes different beads and different binding and buffer chemistry to optimally extract DNA from these sources.

 

In this section, you’ll find KingFisher system-optimized kits, reagents, and protocols that can be used to maximize purity and yield for each extraction target and sample type. The table summarizes the recommended methodology for each type of analyte and application.



Kit, reagent, and protocol selection guide

Use our selection tool to find the right magnetic bead-based kit for your automated sample preparation. MagMAX, Dynabeads, and SMART Digest kits can purify nucleic acids, proteins, cells, exosomes, and virus from a wide range of sample types and are optimized for automated Kingfisher purification systems. 

Go directly to our Kit, Reagent, and Protocol Selection Guide


Downloading protocols

KingFisher BindIt or BindIx software is included free with all KingFisher instruments so that you can create, download, run, modify and store protocols for your applications. The software allows you to define and edit steps, parameters, plates, and reagents for every part of your protocol—bind, wash, and elute.

 

For any MagMAX or Dynabeads kits or reagents in the Kits, Reagents, and Protocols Selection Guide, click View Protocols to review automated protocols for KingFisher instruments. Then follow these steps to download and install the protocols you want.

KingFisher BindIx software (for KingFisher Apex systems)

KingFisher BindIt software (for KingFisher Flex and Duo Prime systems)

  1. Make sure you have BindIt 4.1 Software installed on the computer you use with your KingFisher instrument. Download the latest BindIt Software here.
  2. Find the kit you want in the Kit, Reagent, and Protocol Selection Guide and click View Protocols. You’ll see the product page for that kit.
  3. Scroll to the Protocols section at the bottom and click the protocol you want to download. Make sure it’s for the correct instrument.
  4. Save the protocol on your computer in the preferred folder.
  5. Connect your PC to your KingFisher instrument with a USB cable and select Home > Connect in BindIt Software.
  6. You can start the protocol from the BindIt Software or transfer it onto the instrument. If you transfer the protocol, select Home > Transfer in BindIt Software.
  7. Select Upload and find the saved protocol. Click Open.
  8. Select the folder (category) in which you want to store the protocol on the instrument. Your protocol can be now found in the instrument memory.
  9. Disconnect your instrument from BindIt and check using the keypad that your protocol is in the desired folder.

SARS-CoV-2

KingFisher systems are on the front line of the battle helping labs accelerate RNA extraction throughput to handle the huge demand for fast and accurate SARS-CoV-2 test results. MagMAX viral/pathogen kits on KingFisher instruments provide a sensitive and simple method for nucleic acid extraction for SARS-CoV-2 testing, and protocols have recently been streamlined to further increase throughput.

Sample viral/pathogen analysis workflow

Sample viral/pathogen analysis workflow

Recommended SARS-CoV-2 test kits

When to use MagMAX SARS-CoV-2 viral/pathogen kits

MagMAX viral/pathogen kits are designed for high nucleic acid recovery and are compatible with low-titer samples. There are only two protocols for low- to high-volume input range. No carrier is required, which means less interference with downstream NGS and fewer required steps.

 

MagMAX Viral/Pathogen Nucleic Acid Isolation Kits I and II (MVP I and II) are recommended for use by research labs. Both kits have been validated with newly developed, streamlined protocols allowing a sample input volume of 200 µL (vs standard 400 µL) with half the amount of extraction reagents.

 

The two MagMAX kits are functionally identical, as are the chemistry driving the RNA to the beads and the overall workflow. The two differences are: (a) A raw material change in the bead component due to unprecedented SARS-CoV-2 demand, and (b) The MVP II kit is available only in bulk prep format.

More information on SARS-CoV-2 testing

For application notes and supporting materials about using KingFisher instruments and MagMAX or Dynabeads kits for SARS-CoV-2 testing and isolation, see the Resources tab.

 

For performance data, application notes, and detailed protocol information for any of these kits, follow its Learn More or Cat. No. link above.

 

To view a workflow video for the MagMAX Viral/Pathogen Nucleic Acid Isolation Kits with KingFisher instruments, follow one of its Cat. No. links above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab.


Genomic DNA research

Isolating genomic DNA (gDNA) from different sample types has historically required multiple purification kits. With the MagMAX DNA Multi-Sample Ultra 2.0 Kit or KingFisher Flex Ready DNA Ultra 2.0 Prefilled plates and KingFisher instruments, you can isolate gDNA from 50 µL to 2mL of whole blood, saliva, buffy coat, buccal swabs, or other biological samples. The resulting purified gDNA is ideal for many downstream molecular biology applications such as qPCR, NGS, and microarray analysis.

 

One easily accessible, non-invasive, reliable source of gDNA suitable for downstream genetic analysis is saliva. The MagMAX gDNA Saliva Kit, developed for a leading genomics company for use with KingFisher instruments, is specifically optimized for saliva gDNA isolation.

Sample gDNA workflow

Sample gDNA workflow

Sample gDNA from saliva workflow

Sample gDNA from saliva workflow

Recommended gDNA kits

When to use MagMAX gDNA kits

Use the MagMAX DNA Multi-Sample Ultra 2.0 Kit to recover high-purity gDNA from both small- and large-volume inputs. The kit produces maximum yield by treating samples with proteinase K (PK) and P-enhancer, followed by an innovative lysis/binding solution to simultaneously lyse cells, denature proteins, and capture gDNA on the high-binding magnetic beads. Designed for flexibility, the kit allows you to skip sample and buffer volume normalization steps for trouble-free processing. For example, you can run 50–µL and 400–µL samples at the same time on the same plate.

 

Reagents for the MagMAX DNA Multi-Sample Ultra 2.0 Kit are also available in bulk, allowing you to process 2,000 50–200 µL samples or 1,000 200–400 µL samples.

 

KingFisher Ready DNA Ultra 2.0 Prefilled Plates contain the same reagents and produce the same high-quality results as the MagMAX DNA Multi-Sample Ultra 2.0 Kit, but the plates are already conveniently filled with reagents. This eliminates the need to dispense lysis, wash solutions, and magnetic beads. The protocol is simple: just peel off the foil, add sample and enhancer/Proteinase K solutions, and insert into the instrument.

MagMAX gDNA Saliva Kit

Use the MagMAX gDNA Saliva Kit with KingFisher instruments to recover high-purity gDNA from saliva samples. The kit treats samples with an innovative lysis/binding solution to simultaneously lyse cells, denature proteins, and capture gDNA on the high-binding magnetic beads. Designed for flexibility, the kit allows you to skip sample and buffer volume normalization steps for trouble-free processing. For example, you can run 50-µL and 400-µL samples at the same time on the same plate. The workflow is compatible with saliva collection devices like the Oragene® from Genotek, the GeneFiX™ from IsoHelix, and the SalivaGard® from Biomatrica.

More information on MagMAX gDNA kits

For performance data and protocol information for any of these MagMAX kits, follow its Learn More link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab.

 


Cell-free DNA and RNA

“Liquid biopsies” of cell-free DNA (cfDNA) or total nucleic acids (cfTNA) from plasma, serum, or urine represent a non-invasive approach to detect and monitor targets associated with disease states such as cancer. MagMAX cell-free nucleic acid kits on KingFisher instruments are ideal for liquid biopsy research. They are optimized specifically for enrichment of cell-free DNA (cfDNA) and total nucleic acid (cfTNA), not genomic DNA (gDNA), which means increased recovery and lower starting volumes. 

 

Sample cfDNA or cfTNA liquid biopsy workflow

Sample cfDNA or cfTNA liquid biopsy workflow

Recommended cfDNA and cfTNA kits

When to use MagMAX cfDNA and cfTNA kits

Liquid biopsies most often utilize cell-free DNA (cfDNA) that is derived from both normal and cancerous cells. The tumor-only supply of DNA in the bloodstream is more commonly referred to as circulating tumor DNA (ctDNA), which is loaded with information about a tumor that would otherwise be difficult to access. In these kits, magnetic beads and optimized chemistry combine for more efficient nucleic acid isolation that specifically recovers the fragmented cfDNA while leaving the larger DNA molecules behind. This aspect of enrichment for the cfDNA portion of the total nucleic acid ensures that the shorter, ctDNA is concentrated and ready for downstream analysis using qPCR, digital PCR (dPCR), or next-generation sequencing (NGS).

 

While the importance of cfDNA is becoming established, it may not tell the whole story. cfRNA sequences serve as critical biomarkers in the form of microRNAs and gene fusions. The inclusion of cfRNA with cfDNA in the Total Nucleic Acid Kit provides an opportunity for researchers to expand their studies and advance human health.

 

Use the MagMAX Cell-Free DNA Isolation Kit with KingFisher instruments, Oncomine assays, and other downstream applications to optimize the liquid biopsy workflow from extraction and purification to analysis.

 

Use the MagMAX Cell-Free Total Nucleic Acid Isolation Kit to extend your liquid biopsy workflow to include both cell-free DNA and RNA (cfRNA) from the same sample.

More information on MagMAX cfDNA and cfTNA kits

For performance data, application notes, and detailed protocol information for either of these MagMAX kits, follow its Learn More link above.

 

To view a workflow video for the MagMAX Cell-Free DNA Isolation Kit with KingFisher instruments, follow its Cat. No. link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab.


FFPE DNA and RNA

Clinical cancer research samples are routinely formalin fixed and paraffin embedded (FFPE) for histopathological analysis and tissue archiving. This method preserves the tissue and allows for long-term storage, but the extensive chemical crosslinking of nucleic acids can complicate downstream processing for applications such as next-generation sequencing (NGS).

 

With the MagMAX FFPE DNA/RNA Ultra Kit and KingFisher instruments, you can sequentially isolate DNA, RNA, microRNA, or both DNA and RNA from FFPE and fixed samples. The genomic DNA (gDNA) extracted is high quality with yields comparable to fresh-frozen blood. The kit is ideal for use with Ion Torrent™ Oncomine™ assays as well as PCR and NGS, even when the FFPE tissue is more than 25 years old.

Sample FFPE Isolation workflow

Sample FFPE Isolation workflow

Recommended FFPE DNA and RNA kits

When to use

Use the MagMAX FFPE DNA/RNA Ultra Kit for fast, reliable nucleic acid isolation from FFPE samples. You can even extract DNA and RNA sequentially from the same sample in a single workflow, drawing richer data from your precious samples. For example, you can investigate important biomarkers, such as hotspot mutations, CNVs, gene fusions, and indels.

 

The kit has been validated for multiple research sample types, including solid tumor biopsies, core needle biopsies, and fine-needle aspirates. It is also compatible with CitriSolv™ clearing agent, a deparaffinization alternative to xylene that is less toxic and can be used without a chemical fume hood.

 

To speed up the deparaffinization process, pair the FFPE DNA/RNA Ultra Kit with the Applied Biosystems™ AutoLys™ system. Comprised of pretreated tubes and a rack system to reduce pipetting steps, AutoLys tubes and accessories eliminate the need for hazardous chemicals and minimizes loss of tissue sample.

More information

For performance data, application notes, and detailed protocol information for this MagMAX kit, follow its Learn More link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab.


Total RNA and miRNA

Isolating different types of RNA from different sample types for gene expression studies has historically required multiple purification kits. The MagMAX mirVana Total RNA Isolation Kit is optimized to extract Total RNA, including small RNAs and microRNAs (miRNAs), from most biological samples, including plasma/serum, whole blood, solid tissue, cultured cells, urine, and more. Extraction is phenol-free and non-organic.

Sample total RNA or miRNA workflow

Sample total RNA or miRNA workflow

Recommended Total RNA Isolation Kit

When to use MagMAX mirVana Total RNA Isolation Kit

Use the MagMAX mirVana Total RNA Isolation Kit for low- or high-throughput isolation of true total RNA that covers the full range of RNA types present in cells, from small RNAs such as microRNAs (miRNAs) to non-coding RNA, messenger RNA, and ribosomal RNA. The kit elutes highly concentrated RNA in as little as 20 µL of nuclease-free water and its chemistry stabilizes and preserves RNA over time.

 

The extracted RNA is suitable for a range of downstream gene expression applications such as miRNA-Seq and qRT-PCR. Workflows have been validated for the TaqMan™ Advanced miRNA cDNA Synthesis Kit and other TaqMan Advanced miRNA Assays.

More information on MagMAX mirVana Total RNA Isolation Kit

For performance data for this MagMAX kit, follow its Learn More link above.

 

For protocol information and application notes for this MagMAX kit, or to view a workflow video for this kit (manual isolation only), follow its Cat. No. link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab.


mRNA

Research on transcriptomics requires messenger RNA (mRNA), which can be sourced from a wide variety of samples. Dynabeads mRNA isolation beads specifically capture mRNA molecules from a wide variety of crude samples by exploiting the base pairing between the polyA residues at the 3’ end of most mRNA and the Oligo(dT)25 residues covalently coupled to the surface of the Dynabeads. Automated protocols on KingFisher instruments take only 15 minutes and the isolated mRNA is suitable for use in all downstream applications.

Sample viral/pathogen analysis workflow

Sample mRNA research workflow

Recommended kits for mRNA research

When to use Dynabeads mRNA Purification Kits

The Dynabeads mRNA Purification Kit takes Total RNA as input and is a good choice when you have already isolated Total RNA, such as from the MagMAX mirVana Total RNA Isolation Kit.

 

However, only 1–5% of the Total RNA content of cells is actually mRNA. If all you need is the mRNA transcriptome, there is no need to isolate Total RNA. Dynabeads mRNA DIRECT™ purification kits and technologies target only the mRNA molecules from a wide variety of crude samples and pull them directly out of solution. Ribosomal RNA, DNA, proteins, and small RNA molecules (such as transfer RNA, micro RNA, and small nucleolar RNA) do not bind to the beads and are discarded.

Yield from a Total RNA Prep

With all three kits, you can synthesize complementary DNA (cDNA) directly on the beads to create a reusable solid-phase cDNA library, and even perform qRT-PCR directly on the bead-bound cDNA for sensitive quantitative analysis of the entire transcriptome, down to the level of a single isolated cell.Use the Dynabeads mRNA Purification Kit to isolate mRNA when you have already purified Total RNA.

 

Use the Dynabeads mRNA DIRECT Purification Kit to isolate mRNA directly from the crude lysate of nearly any biological sample of vertebrate, invertebrate, plant, fungal, or bacterial origin. The sensitive technology enables cDNA synthesis and cDNA library construction from starting samples as small as a single cell.

 

The Dynabeads mRNA DIRECT Micro Purification Kit handes the same applications as the mRNA DIRECT Purification Kit but is optimized for micro-size samples of 10,000 cells, 5 mg tissue, or less.

More information on Dynabeads kits

For protocols for any of these Dynabeads kits, follow its Cat, No. link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab.


Viral/pathogen DNA and RNA

The detection of viruses in human biofluids is crucial to infectious disease research and management. Viral load testing is used to assess the efficacy of antiviral therapy, as well as to monitor disease progression and uncover the presence of an infection at an earlier stage. MagMAX viral/pathogen kits on KingFisher instruments provide a sensitive and simple method for nucleic acid extraction from samples containing viruses or other pathogens.

 

Note: For specific information about using MagMAX Viral/Pathogen kits for SARS-CoV-2 testing, see SARS-CoV-2 testing above.

Sample viral/pathogen analysis workflow

Sample viral pathogen analysis workflow

Recommended viral/pathogen test kits

When to use MagMAX viral/pathogen kits

MagMAX viral/pathogen kits are designed for high nucleic acid recovery and are compatible with low-titer samples. There are only two protocols for low- to high-volume input range. No carrier is required, which means less interference with downstream NGS and fewer required steps.

 

The MagMAX Viral/Pathogen Nucleic Acid Isolation Kits provide a sensitive and simple method for RNA and DNA extraction from samples containing viruses and Gram-negative bacteria, as demonstrated by successful detection of as few as 50 copies of input into multiple sample types.

 

The MagMAX Viral/Pathogen Ultra Nucleic Acid Isolation Kit (A42356) contains the same reagents as the standard Viral/Pathogen Nucleic Acid Isolation Kits, as well as an extra enzyme mix needed to lyse and recover nucleic acids from difficult pathogens like Gram-positive bacteria, yeast, and fungus. It can be used for broadbased detection and identification of unknown pathogens, including difficult-to-detect respiratory tract microbiota. Because the extra enzymes are not needed for viral RNA isolation, this kit is not recommended for SARS-CoV-2 testing.

 

Note: If you need to isolate SARS-CoV-2 RNA, make sure to order the MagMAX Viral/Pathogen II (MVPII) Nucleic Acid Isolation Kit (Cat. No. A48383R), not the MagMAX Viral/Pathogen Ultra Nucleic Acid Isolation Kit (Cat. No. A42356). The Ultra version contains an extra enzyme mix needed to lyse Gram-positive bacteria and fungus, which is not needed for SARS-CoV-2 RNA isolation.

More information about MagMAX viral/pathogen kits

For performance data, application notes, and detailed protocol information for any of these MagMAX kits, follow its Learn More or Cat. No. link above.

 

To view a workflow video for the MagMAX Viral/Pathogen Nucleic Acid Isolation Kits with KingFisher instruments, follow one of its Cat. No. links above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the  Accessories tab.


Microbiome research with DNA and RNA

The human microbiome has emerged as a key player in the diagnosis and treatment of disease, but identifying and quantifying the microbes that make it up constitute a major challenge. Although metagenomics sequencing technology can providing a comprehensive profile of all the genes in an environmental sample, it requires an adequate sample of pure, high-quality DNA and RNA. The MagMAX Microbiome Ultra Nucleic Acid Isolation Kit and KingFisher instruments can provide that sample in an efficient, automated workflow.

Sample microbiome research workflow

Sample microbiome research workflow

Recommended microbiome research kit

When to use MagMAX Microbiome Ultra Kit

The human microbiome is complex and multifaceted, and each body habitat harbors different dominant signature taxa. The gut contains the bulk of microbiota associated with the human body and has been one of its most thoroughly examined ecosystems. Fecal samples are commonly used for these analyses because they are easy to obtain and contain a large amount of biomass. Body fluids like saliva and urine are less studied, but each contains unique bacterial communities.

 

Use the MagMAX Microbiome Ultra Nucleic Acid Isolation Kit to isolate the total nucleic acid from feces, urine, saliva, and other samples. It can extract high-yield, high-purity, high-quality, inhibitor-free DNA and RNA that is ideal for metagenomics research and other downstream applications. Enzyme-based digestion allows recovery of nucleic acids from a wide range of microorganisms without additional processing steps.

 

The kit can isolate both viral and bacterial nucleic acids, and has been tested with mammalian, reptilian, fish, and bird fecal samples; VTM, skin, and milk samples; and yeast and bacterial cultures. It is recommended for research labs that are developing their own microbiome tests and need low- and/or high-throughput solutions for isolating total nucleic acid.

More information about MagMAX viral/pathogen kits

For performance data and application notes for this MagMAX kit, follow its Learn More link above.

 

For protocol information for this MagMAX kit, follow one of its Cat. No. links above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab


Plant DNA and RNA

Extracting DNA and RNA from plants brings unique challenges in dealing with carbohydrates, phenolics, and other compounds abundant in plant tissues. MagMAX™ Plant DNA Isolation Kit, automated with KingFisher instruments, can isolate nucleic acids from a wide variety of plant species and tissue types.

Sample plant DNA workflow

Sample plant DNA workflow

Sample plant RNA workflow

Sample plant RNA workflow

Recommended Plant DNA and RNA extraction kits

When to use MagMAX Plant DNA or RNA kits

Plant cell walls can be very difficult to disrupt and lysates often contain significant amounts of compounds such as tannins, phenolics, and complex polysaccharides that can affect nucleic acid quality and inhibit downstream reactions. Despite these challenges, these MagMAX kits can effectively isolate DNA or RNA from diverse plant samples such as Arabidopsis thaliana, bell pepper leaves, canola/rapeseed leaves, corn leaves, green onion, lemon leaves, oat leaves, orange, pine needles, spinach leaves, tomato leaves, and wheatgrass leaves.

 

Use the MagMAX Plant DNA and RNA Kits with KingFisher instruments for automated purification of DNA and RNA from a wide variety of plant species and tissue types, including plant fungus. Purified DNA or RNA is free of proteins, nucleases, and other contaminants, inhibitors, and nucleic acids, and can be used in a wide range of downstream applications such as PCR, qPCR, and NGS. The kits are optimized for isolation of DNA and RNA from 10–100 mg of plant samples.

More information on MagMAX Plant DNA and RNA kits

For performance data and protocols for either of these MagMAX kits, follow its Cat. No. link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab


Immunoprecipitation

Immunoprecipitation (IP) or co-immunoprecipitation (co-IP) can be used to isolate and concentrate a particular protein or protein complex from a sample containing many thousands of different proteins. Dynabeads magnetic beads, which maximize yield and reproducibility and minimize nonspecific binding, have become the gold standard for magnetic bead-based IP. With automation protocols now available for KingFisher instruments, you can be ready to detect and analyze proteins in just 40 minutes with minimal hands-on time.

Sample immunoprecipitation (IP) workflow

Sample immunoprecipitation (IP) workflow

Recommended kits and products for immunoprecipitation

When to use Dynabeads for immunoprecipitation

There are two variations on the IP process. If your target protein is abundant, you can use the direct method, in which the primary antibody (Ab) is first incubated with the Dynabeads magnetic beads, followed by a wash to remove unbound antibodies. The Ab-bound beads are then incubated with with the sample for capture, followed by washing and elution.

 

If the target protein is not abundant, you can use the indirect method, in which the primary Ab is first incubated with the cell lysate to form an Ab-antigen complex. The complex is then captured by adding Dynabeads beads to the sample, followed by washing and elution.

 

Although IP can be performed manually, the number of washing steps and the hands-on time increase proportionally with the number of samples. Automating the process with KingFisher instruments can eliminate inconsistencies introduced by pipetting and other manual handling. The automated protocols replicate the manual protocols, obtaining equally high target protein yield and the same low non-specific binding and high reproducibility. Whether you are working with 10 or 96 samples, the IP protocol takes less than 40 minutes. Just load the reagents on the plates, push the Start button, and by the time you have prepared for downstream analysis, the IP is done. Some optimization, such as incubation time, might be necessary depending on your Ab and the abundance and/or specificity of your target protein.

 

Deciding among the kits depends on the target protein you need to isolate. If you have a primary antibody to the protein, we recommend using Dynabeads Protein A or G or their associated kits. If you have a secondary antibody, use one of the Dynabeads M-280 IgG products. Finally, if you have a biotinylated antibody or ligand, use Dynabeads M-280 Streptavidin. 

More information on Dynabeads and immunoprecipitation

For performance data, application notes, and protocol information for any of these Dynabeads products, follow its Cat. No. link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab


Peptide mapping and quantitation

Peptide mapping is a critical workflow in biotherapeutic protein characterization and is essential for elucidating the primary amino acid structure of proteins. In order to generate a peptide map, the therapeutic protein must first be digested into its constituent peptides via a chemical or enzymatic reaction. But methods currently employed for digestion and mapping involve lengthy, multi-step, manual processes subject to low levels of reproducibility, sensitivity, and data confidence.

 

Thermo Scientific SMART Digest magnetic kits paired with KingFisher instruments can shorten a 24-hour process to an hour or less, automating much of the hands-on work and eliminating sources of variation and manual error.

Sample peptide mapping workflow

Sample Peptide mapping workflow

Sample peptide quantitation workflow

sample peptide quantitation workflow

Recommended peptide mapping and quantitation kits

When to use SMART Digest kits

Thermo Scientific SMART Digest Kits paired with KingFisher instruments represent the latest advance in automated protein digestion for peptide mapping and quantitation applications. The kits make protein digestion fast, simple, reproducible, and easy to automate.

 

The protein digestion kits use optimized heat-stable enzymes on magnetic beads, minimizing the possibility of post-translational modifications. Automating the process minimizes manual handling, ensures the optimal timing of reactions, and reduces high-temperature exposure, increasing reproducibility of results.[1]

 

Immunoaffinity (IA) kits couple enzymatic digestion with IA capture on a single bead for high sensitivity. This innovation generates high confidence in your high-performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometry (LC/MS) results. SMART Digest Kits especially benefit biomarker and biopharmaceutical characterization and quantitation applications.

 

For peptide mapping applications, choose among optimized heat-stable trypsin, chymotrypsin, or proteinase K enzymatic digestion for sequence-specific cleavage of your protein. The enzyme, on magnetic beads, is activated at elevated temperatures for accelerated digestion under protein denaturing conditions.

  • Trypsin cleaves the C-terminal side of lysine and arginine amino acid residues
  • Chymotrypsin cleaves the peptide bonds formed by aromatic residues such as tyrosine, phenylalanine, and tryptophan
  • Proteinase K cleaves the peptide bond adjacent to the carboxyl group of aliphatic and aromatic amino acids with blocked alpha amino groups

 

For peptide quantitation applications, SMART Digest IA Kits perform trypsin digestion with immunoaffinity capture in a single well. The immunoaffinity moiety (streptavidin, protein A, or protein G) is typically employed to increase sensitivity by purifying your low-level proteins from complex biological matrices.

  1. Farrell A, Bones J, Cook K, Patel S, Schwahn A, Bardsley J. High-precision, automated peptide mapping of proteins. Thermo Fisher Scientific Application Note 21682. Full text

More information on SMART Digest products

For performance data and application notes for any of these SMART Digest products, follow its Cat. No. link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab.


Cells and exosomes

Above the molecular level of nucleic acids and proteins, Dynabeads magnetic beads reagents can be used with KingFisher instruments for automated isolation of cells and exosomes. For example, T cells can be isolated, activated, and expanded using Dynabeads reagents for use in downstream applications such as cytotoxicity or gene editing. Exosomes and circulating tumor cells (CTCs) can be isolated and analyzed for liquid biopsies. The use of Dynabeads reagents helps ensure high yields of pure, viable, and functional cells whose native characteristics are preserved, while KingFisher system automation reduces hands-on time and improves reproducibility.

Recommended cell and exosome research kits

When to use Dynabeads for cell and exosome isolation

There are three variations on the cell isolation process. In positive cell isolation, Dynabeads reagents bind the target cells and the bead/cell complexes are pulled to the KingFisher magnetic tips. After discarding the supernatant, only your pure target cells remain.

 

In negative cell isolation, Dynabeads Untouched™ kits are used to deplete unwanted cells, leaving bead-free and untouched target cells in the sample. For example, the Dynabeads Untouched Human T Cells Kit includes an antibody mixture to remove B cells, natural killer (NK) cells, monocytes, platelets, dendritic cells, granulocytes, and erythrocytes, leaving all T cells (and only T cells) for further use. Other Dynabeads Untouched kits can remove all but the CD4 or CD8 T-cell subsets.

 

If you are interested in multiple cell types, you can use cell-specific depletion Dynabeads reagents to remove all unwanted cell types from a heterogeneous sample with >99% efficiency. Only cells of interest will be left in the depleted supernatant.

Dynabeads T-cell activation products mimic the in vivo interaction of T cells with APCs by conjugating the two APC activation signals (CD3 and CD28) to magnetic beads.

Dynabeads T cell activation products

Isolation of T cells and subsets is often followed by activation and expansion. Dynabeads CD3/CD28 technology mimics in vivo T-cell activation via antigen-presenting cells (APCs), and allows gentle and efficient physiological activation and expansion of both mouse and human T cells.

 

To isolate and enrich total exosomes from cell culture, use the Total Exosome Isolation reagents. You can then further purify them into specific subpopulations using Dynabeads CD9, CD63, CD81, or EpCam-specific reagents. Or, combine the Dynabeads Straptavidin reagent with your choise of a biotinylated antibody to purify any population based on a surface antigen.

 

The downstream application determines the number of magnetic beads to use for exosome capture. For flow cytometry, keep the number of beads low to ensure a high density of exosomes per bead and obtain a strong signal. For western blotting, use a large number of beads to maximize total surface area and give a strong signal. This will leave very few exosomes in the supernatant. 

More information on Dynabeads for cell and exosome isolation

For performance data, application notes, and protocol information for any of these Dynabeads products, follow its Learn More link above.

 

For information about plastics and plate setup products to use with KingFisher instruments, see the Accessories tab.