Shop All Protein Analysis Kits

Qubit™ Protein Assay Kit (Invitrogen™)

The Qubit Protein Assay Kit is designed specifically for use with the Qubit Fluorometer. Using between 1 and 20 µl of your sample, this assay can quantitate samples ranging from 12.5 µg⁄ml to 5 mg⁄ml and exhibits low protein to protein variation. The assay is highly selective for proteins and is designed to be accurate in the presence of reducing reagents, but not in the presence of a large amount of detergent. Common contaminants, such as reducing reagents (DTT, β-mercaptoethanol), salts, free nucleotides, amino acids, solvents, or DNA, but not detergents, are well tolerated in the assay. Slight protocol modifications are required for other contaminants. The kit provides concentrated assay reagent, dilution buffer, and pre-diluted BSA standards. Simply dilute the reagent using the buffer provided, add your sample (any volume between 1 µL and 20 µL is acceptable), and read the concentration.

Which product to choose for fluorometric protein quantitation?
• For 1–20 samples: use this Qubit Protein Assay Kit with the Qubit Fluorometer
• For 20–2000 samples: use the Quant-iT Protein Assay Kit with microplate reader

Notes:
1. All Qubit assay kits can be used with the Qubit 1.0, Qubit 2.0, Qubit 3, and Qubit 4 fluorometers.
2. 500 µL thin-walled PCR tubes are required but not included.

Pro-Detect™ Rapid Streptag II Competitive Assay Kit (Thermo Scientific™)

The Thermo Scientific Pro-Detect Rapid Streptag II Competitive Assay Kit is a single-use, dipstick lateral flow kit for quick, easy detection of Strep Tag II (sequence: PQKWSHPQFEK)-tagged proteins in protein expression and purification systems. This lateral flow assay uses membrane-based strips coated with dye-bound conjugated antibodies specific to Streptag II tag to provide a qualitative detection within ten minutes. This assay can be used to detect Streptag II-tagged proteins directly from cell culture media (supernatant) or cell lysates without any special instrumentation or handling.

Features of the Pro-Detect Rapid competitive assay kits:
Easy to adopt—lateral flow strips can be used early in and throughout expression to monitor and confirm expression of desired protein-tagged target
Simple, fast method—simply dilute sample using provided diluent and dip in the lateral flow strip for qualitative results within 5-10 minutes
Sensitive—detect protein tags in supernatant or cell lysate samples down to low µg/mL concentrations
Specific—assay does not cross-react with common lysis and protein preparation reagents
Reliable—results are similar to other antibody immunoassays, including western blot and ELISA
Long shelf life—stable for at least one year at 4°C

Each Pro-Detect Rapid competitive assay is performed by simply adding a properly diluted tissue culture supernatant, lysate, or purified protein sample to a test tube or microplate well and dipping in a lateral flow strip. Gold-conjugated detection antibodies embedded in the strip form complexes with the tagged protein in the sample. In the presence of target protein tag, the dye-bound conjugated antibodies will bind to the tagged protein present in the sample instead of the protein tag bound at the test lines. Therefore, in a positive result, a competitive lateral flow assay will show decreasing intensity of the visible test lines until all test lines have disappeared. When there is no target protein tag present, the dye-bound conjugated antibodies specific to the target will bind at the test line(s), resulting in visual test lines in a negative result.

ProQuest™ Two-Hybrid System with Gateway™ Technology (Invitrogen™)

The ProQuest™ Two-Hybrid System with Gateway® Technology is an in vivo yeast-based system for identifying interactions between two proteins. This interaction reconstitutes a functional transcription factor that activates chromosomally integrated reporter genes where transcriptional activation is monitored by the growth of cells on selective media.
The ProQuest™ System features:
• Gateway® Technology to allow rapid and easy generation of bait and prey constructs, and to facilitate downstream application
• Low copy-number DBD (DNA Binding Domain) and AD (Activation Domain) vectors (ARS/CEN) to control over-expression and increase reproducibility
• Three different reporter genes (HIS3, URA3, and lacZ) with independent promoter regions to rapidly weed out false positives. URA3 reporter gene allows for both positive and negative selection, enabling advanced two-hybrid techniques such as reverse two-hybrid.

• A set of strong-to-weak Gateway®-based control interactions to evaluate results and assess the strength of the interaction being tested

Pro-Detect™ Rapid AVI Competitive Assay Kit (Thermo Scientific™)

The Thermo Scientific Pro-Detect Rapid AVI Competitive Assay Kit is a single-use, dipstick lateral flow kit for quick, easy detection of AVI (sequence: GLNDIFEAQKIEWHE)-tagged proteins in protein expression and purification systems. This lateral flow assay uses membrane-based strips coated with dye-bound conjugated antibodies specific to AVI tag to provide a qualitative detection within ten minutes. This assay can be used to detect AVI-tagged proteins directly from cell culture media (supernatant) or cell lysates without any special instrumentation or handling.

Features of the Pro-Detect Rapid competitive assay kits:
Easy to adopt—lateral flow strips can be used early in and throughout expression to monitor and confirm expression of desired protein-tagged target
Simple, fast method—simply dilute sample using provided diluent and dip in the lateral flow strip for qualitative results within 5-10 minutes
Sensitive—detect protein tags in supernatant or cell lysate samples down to low µg/mL concentrations
Specific—assay does not cross-react with common lysis and protein preparation reagents
Reliable—results are similar to other antibody immunoassays, including western blot and ELISA
Long shelf life—stable for at least one year at 4°C

Each Pro-Detect Rapid competitive assay is performed by simply adding a properly diluted tissue culture supernatant, lysate, or purified protein sample to a test tube or microplate well and dipping in a lateral flow strip. Gold-conjugated detection antibodies embedded in the strip form complexes with the tagged protein in the sample. In the presence of target protein tag, the dye-bound conjugated antibodies will bind to the tagged protein present in the sample instead of the protein tag bound at the test lines. Therefore, in a positive result, a competitive lateral flow assay will show decreasing intensity of the visible test lines until all test lines have disappeared. When there is no target protein tag present, the dye-bound conjugated antibodies specific to the target will bind at the test line(s), resulting in visual test lines in a negative result.

Pierce™ c-Myc-Tag Magnetic IP/Co-IP Kit (Thermo Scientific™)

The Thermo Scientific Pierce Magnetic c-Myc-Tag IP/Co-IP Kit contains specific immunoaffinity magnetic beads and reagents to perform immunoprecipitation assays of c-Myc fusion proteins or co-IP experiments using c-Myc-tagged bait proteins.

Unlike traditional IP procedures based on capture with Protein A/G beads, this kit uses magnetic beads containing pre-immobilized anti-c-Myc antibody. These Pierce Anti-c-Myc Magnetic Beads ensure specific binding of c-Myc tagged protein complexes from biological samples. Because the antibody is covalently attached to the beads, IP-targets are easily eluted and recovered without antibody contamination. The complete IP kit includes the magnetic beads, lysis/wash buffer, low-pH elution buffer, neutralization buffer, c-Myc-tag positive control lysate, and non-reducing sample buffer for SDS-PAGE. Protocols are provided for both manual and automated magnetic separation workflows. Sufficient components are provided to perform 40 IP or co-IP assays.

Features of the c-Myc-Tag Magnetic IP/Co-IP Kit:

Specific magnetic beads—covalently immobilized high-quality anti-c-Myc monoclonal antibody enables high yields of immunoprecipitation products
Low non-specific binding—stable, pre-blocked beads and specific antibody minimize off-target binding
Trouble-free elution—low-pH elution buffer ensures recovery of c-Myc-tagged protein interaction complexes without antibody leaching contamination
Convenient and fast—complete kit and easy-to-follow instructions provide optimized protocols to perform IP or Co-IP experiment in approximately 1 hour
Versatile—magnetic beads are compatible with manual and automated magnetic separation workflows (e.g., Thermo Scientific KingFisher Instruments)

The c-Myc peptide (EQKLISEEDL) derived from the C-terminus region of human c-Myc protein is one of several fusion protein tags used for recombinant protein expression. The Pierce c-Myc Magnetic IP/Co-IP Kit uses a specific, high-affinity immobilized antibody (clone 9E10) for rapid immunoprecipitation of c-Myc tagged fusion proteins from bacterial and mammalian cell lysates, as well as from lysates prepared with the Pierce Human in vitro Translation Kits. The beads are incubated with a cell lysate containing c-Myc tagged protein, the fusion protein is captured, and the beads are subsequently washed and then eluted using low-pH elution buffer or non-reducing sample buffer. The protocol and buffers have been optimized for both IP and co-IP reactions, enriching for specific protein interaction complexes in the eluted samples. Anti-c-Myc antibody can be used to detect c-Myc tagged protein by Western blot analysis.

For information about the binding capacity and other properties of the magnetic beads used in this kit, see Pierce Anti-c-Myc Magnetic Beads (Part No. 88842).

Related Products
Pierce™ c-Myc-Tag IP/Co-IP Kit

Pro-Detect™ Rapid Myc Competitive Assay Kit (Thermo Scientific™)

The Thermo Scientific Pro-Detect Rapid Myc Competitive Assay Kit is a single-use, dipstick lateral flow kit for quick, easy detection of Myc (sequence: EQKLISEEDL)-tagged proteins in protein expression and purification systems. This lateral flow assay uses membrane-based strips coated with dye-bound conjugated antibodies specific to Myc tag to provide a qualitative detection within ten minutes. This assay can be used to detect Myc-tagged proteins directly from cell culture media (supernatant) or cell lysates without any special instrumentation or handling.

Features of the Pro-Detect Rapid competitive assay kits:
Easy to adopt—lateral flow strips can be used early in and throughout expression to monitor and confirm expression of desired protein-tagged target
Simple, fast method—simply dilute sample using provided diluent and dip in the lateral flow strip for qualitative results within 5-10 minutes
Sensitive—detect protein tags in supernatant or cell lysate samples down to low µg/mL concentrations
Specific—assay does not cross-react with common lysis and protein preparation reagents
Reliable—results are similar to other antibody immunoassays, including western blot and ELISA
Long shelf life—stable for at least one year at 4°C

Each Pro-Detect Rapid competitive assay is performed by simply adding a properly diluted tissue culture supernatant, lysate, or purified protein sample to a test tube or microplate well and dipping in a lateral flow strip. Gold-conjugated detection antibodies embedded in the strip form complexes with the tagged protein in the sample. In the presence of target protein tag, the dye-bound conjugated antibodies will bind to the tagged protein present in the sample instead of the protein tag bound at the test lines. Therefore, in a positive result, a competitive lateral flow assay will show decreasing intensity of the visible test lines until all test lines have disappeared. When there is no target protein tag present, the dye-bound conjugated antibodies specific to the target will bind at the test line(s), resulting in visual test lines in a negative result.

Compat-Able™ BCA Protein Assay Kit (Thermo Scientific™)

The Thermo Scientific Compat-Able BCA Protein Assay Kit eliminates interfering substances from protein assay samples to make them compatible with BCA and Bradford assays.

The Compat-Able Reagents remove salts, detergents, reducing agents and other substances from protein samples to eliminate interference with protein assays. The protocol selectively precipitates all of the protein, thereby allowing the non-protein components to be easily decanted and removed. The purified, precipitated protein is redissolved in water, making it ready for accurate determination of the protein concentration by standard protein assay methods. The procedure is accomplished in less than 10 minutes with just a few simple steps. The Compat-Able Preparation Set is available individually or conveniently bundled with either the Thermo Scientific Pierce BCA Protein Assay Kit or the Thermo Scientific Pierce Coomassie Plus Protein Assay Kit.

Features of the Compat-Able Protein Assay Preparation Reagent Kit:

Eliminate incompatibilities—removes nearly any substance that interferes with colorimetric protein assays: salts, detergents, denaturants, reducing agents, and even IEF sample buffer
Simple and fast—prepare samples in less than 10 minutes with four simple steps
Convenient—reagent set is easier to use than homemade TCA or acetone precipitation reagents, thereby producing more consistent results
Scalable—microcentrifuge method is easily scaled to larger centrifuge tube volumes for special needs
Validated—tested and offered together with BCA and Coomassie Plus Protein Assay Kits; protocol integrates control and normalization procedures

Related Products
Compat-Able™ Protein Assay Preparation Reagent Kit
Compat-Able™ Coomassie Plus Protein Assay Kit
Ampule Breakers

Pro-Detect™ Rapid Antibody Isotyping Assay Kit, mouse (Thermo Scientific™)

The Thermo Scientific Pro-Detect Rapid Antibody Isotyping Assay Kit - Mouse is a single-use, dipstick lateral flow kit for quick, easy determination of mouse monoclonal antibody class and subclass identity. This antibody isotyping assay uses membrane-based strips coated with dye-bound conjugated antibodies that provide a visual, color readout of the monoclonal antibody isotype within ten minutes. The kit determines mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM, as well as light-chain identity (kappa vs. lambda light chains).

Features of the Pro-Detect Rapid Antibody Isotyping Assay Kit:
Long shelf life—stable for at least one year at 4°C
Simple workflow—dilute antibody sample using provided diluent and dip in the lateral flow strip
Fast—within 5–10 minutes a band appears that indicates the antibody isotype
Sensitive—detect antibodies in tissue culture media or ascites fluid samples down to low ng/mL concentrations
Specific—assay does not cross-react with fetal bovine serum (FBS)
Reliable—results are similar to standard ELISA-based isotyping assays

Each Pro-Detect Rapid Antibody Isotyping assay is performed by simply adding a properly diluted tissue culture supernatant or mouse ascites sample to a test tube or microplate well and dipping in a lateral flow strip. Gold conjugates embedded in the strip membrane form specific class- and subclass-soluble complexes with the antibodies in the sample. These complexes travel the length of the membrane and are resolved on the anti-isotype and class-specific antibody-impregnated regions of the membrane. Results are displayed as a red band above the printed class or sub-class description, indicating the antibody isotype present.

Determining the class and subclass of a monoclonal antibody is useful in planning the best immunoglobulin purification method. For example, mouse IgA and IgM are best purified by size (i.e., gel exclusion) or using immunoaffinity separation columns. Mouse IgG2a and IgG2b are purified with immobilized Protein A at pH 7–8, while mouse IgG1 binds best to Protein A at pH 8–9. Immunoglobulin that contains kappa light chains can be purified using immobilized Protein L.

The Pro-Detect Rapid Antibody Isotyping Assay Kit is compatible with both tissue culture supernatant and mouse ascites fluid, and is more sensitive than conventional latex bead-based dipstick assays and much faster than ELISA-based isotyping assays. The kit requires only 0.5 µL ascites fluid, 2–22 µL of cell culture supernatant or 1.5 ng of purified antibody.

Pierce™ LAL Chromogenic Endotoxin Quantitation Kit (Thermo Scientific™)

The Thermo Scientific Pierce LAL Chromogenic Endotoxin Quantitation Kit measures the amount of endotoxin in a protein, peptide or antibody sample using the Limulus Amebocyte Lysate (LAL) assay.

Features of the LAL Chromogenic Endotoxin Quantitation Kit:

Sensitive—detect as little as 0.1 EU/mL (approx. 0.01ng endotoxin per mL)
Fast—perform this assay in 30 minutes using a 96-well microplate
AccurateE. coli O111:B4 standard in each kit enables accurate endotoxin quantitation
Versatile—405nm absorbance reading is compatible with common ELISA plate readers

The endotoxin concentration in a sample is measured using the Pierce LAL Chromogenic Endotoxin Quantitation Kit via a chromogenic signal generated in the presence of endotoxins. Samples can be measured on a microplate absorbance reader at 405nm. A standard curve is created using the E. coli endotoxin standard included with each kit to calculate endotoxin levels as low as 0.1 EU/mL, where one endotoxin unit/mL (EU/mL) equals approximately 0.1ng endotoxin/mL of solution. Protein and antibody samples can be assayed in about 30 minutes. Determining endotoxin levels is important to assess the efficiency of endotoxin removal methods and prevent endotoxic shock, inflammation and/or sepsis in tissue culture cells and animals injected with endotoxin contaminated proteins.

Includes:
Kit contains Limulus Amebocyte Lysate (LAL), E. coli endotoxin standard, chromogenic substrate and endotoxin-free water to prepare assay reagents.

Applications:
Quantitation of endotoxin levels in a protein, peptide or antibody solution

The LAL method for measuring endotoxin is based on the interaction of endotoxins with the proenzyme Factor C found in circulating amebocytes of the horseshoe crab Limulus polyphemus. The proteolytic activity of this proenzyme is activated in the presence of lipopolysaccharides (endotoxins) derived from the outer cell membrane of gram-negative bacteria such as E. coli. The Chromogenic Limulus Amebocyte Lysate assay measures endotoxin levels by measuring the activity of this protease in the presence of a synthetic peptide substrate that releases p-nitroaniline (pNA) after proteolysis, producing a yellow color that can be measured by reading the absorbance at 405nm.

To accurately measure endotoxin levels in a sample, the LAL assay uses an endotoxin standard of known concentration that is derived from E. coli strain O111:B4. This standard is provided with each kit and is used to create a standard curve. The endotoxin concentration is determined by extrapolating the absorbance of an unknown sample against this standard curve, similar to ELISA or total protein quantitation assays.

More Product Data
Removing endotoxins using a spin-column format
Eliminate endotoxins from protein and antibody samples

Pierce™ Peroxide Assay Reagent C (Thermo Scientific™)

Thermo Scientific Pierce Peroxide Assay Reagent C is a formulation consisting of 125µM xylenol orange in methanol with BHT. This formulation is one of the component of Quantitative Peroxide Assay Kit used for detecting and measuring hydrogen peroxide levels based on the oxidation of ferrous to ferric ion in the presence of xylenol orange, present in the biological and other liquid samples.

Features of Peroxide Assay Reagent B:

Convenient—no proprietary reagents involved and remove the variability, difficulty and added expense of formulating the reagents yourself
Lipid-compatible formulations— the lipid-compatible formulation provides the best results with lipid-containing samples without first extracting the lipids;however, because this kit does not contain sorbitol, the assay is less sensitive.

Applications of Pierce Quantitative Peroxide Assay Kits:
• General hydrogen peroxide (H2O2) measurement in protein samples
• Quantitation of peroxides in detergents
• Assessment of the effects of peroxides on cellular activity
• Determination of protein glycation; Measurement of peroxide accumulation in lipids

In these assay kits, hydroperoxides convert the Fe2+ to Fe3+ at acidic pH. With the aqueous-compatible formulation, peroxide first reacts with sorbitol, converting it to a peroxyl radical, which in turn initiates Fe2+ oxidation to Fe3+. In the lipid compatible formulation, the peroxide converts the Fe2+ to Fe3+ directly. In a sulfuric acid solution, the Fe3+ complexes with the xylenol orange (XO) dye to yield a purple product having an absorbance maximum at 560nm. Peroxide levels in test samples can be determined by calculation from the know extinction coefficient of the XO-Fe complex or by reference to a standard curve prepared with hydrogen peroxide solution.Most proteins do not interfere with these assays, although some metal chelators may require use of a blank (i.e., control) to mitigate these effects.


Related Products
Pierce™ Quantitative Peroxide Assay Kit (Aqueous)
Pierce™ Quantitative Peroxide Assay Kit (Lipid)
Pierce™ Peroxide Assay Reagent A
Pierce™ Peroxide Assay Reagent B

Pro-Detect™ Rapid V5 Competitive Assay Kit (Thermo Scientific™)

The Thermo Scientific Pro-Detect Rapid V5 Competitive Assay Kit is a single-use, dipstick lateral flow kit for quick, easy detection of V5-tagged proteins in protein expression and purification systems. This lateral flow assay uses membrane-based strips coated with dye-bound conjugated antibodies specific to V5 tag to provide a qualitative detection within ten minutes. This assay can be used to detect V5-tagged proteins directly from cell culture media (supernatant) or cell lysates without any special instrumentation or handling.

Features of the Pro-Detect Rapid competitive assay kits:
Easy to adopt—lateral flow strips can be used early in and throughout expression to monitor and confirm expression of desired protein-tagged target
Simple, fast method—simply dilute sample using provided diluent and dip in the lateral flow strip for qualitative results within 5-10 minutes
Sensitive—detect protein tags in supernatant or cell lysate samples down to low µg/mL concentrations
Specific—assay does not cross-react with common lysis and protein preparation reagents
Reliable—results are similar to other antibody immunoassays, including western blot and ELISA
Long shelf life—stable for at least one year at 4°C

Each Pro-Detect Rapid competitive assay is performed by simply adding a properly diluted tissue culture supernatant, lysate, or purified protein sample to a test tube or microplate well and dipping in a lateral flow strip. Gold-conjugated detection antibodies embedded in the strip form complexes with the tagged protein in the sample. In the presence of target protein tag, the dye-bound conjugated antibodies will bind to the tagged protein present in the sample instead of the protein tag bound at the test lines. Therefore, in a positive result, a competitive lateral flow assay will show decreasing intensity of the visible test lines until all test lines have disappeared. When there is no target protein tag present, the dye-bound conjugated antibodies specific to the target will bind at the test line(s), resulting in visual test lines in a negative result.

GlycoLink™ IP Kit (Thermo Scientific™)

The Thermo Scientific GlycoLink IP Kit provides components for effective immunoprecipitation (IP and co-IP) based on small-scale, covalent, affinity-resin immobilization of antibodies and other glycoproteins via oxidized sugar groups.

Features of the GlycoLink IP Kit:

Efficient immobilization—couple 2 to 10 µg of oxidized antibody or other glycoprotein to 20 µL of resin in 2 hours or less
Stable immobilization—resonance structure of the hydrazone bonds are sufficiently stable so the antibody remains coupled during elution
Preserves binding function—immobilizes IgG via the Fc region, thereby keeping both antigen binding sites available for capturing target
Versatile applications—immobilize any molecule that contains oxidizable sugars, including glycoproteins for use in purifying protein interaction binding partners

The GlycoLink Kit uses hydrazide-activated UltraLink Resin to immobilize and perform 25 IP or Co-IP assays with different antibodies or glycoprotein ligands. The method is ideal for polyclonal antibodies, which typically have abundant carbohydrates (glycosylation) on their Fc portions. Monoclonal antibodies that contain adequate carbohydrates are also effective. Antibodies immobilized by this hydrazide-to-carbohydrate method have unobstructed antigen-binding sites and optimal purification capability. The prepared glycoprotein or antibody columns can be regenerated and reused at least five times for immunoprecipitation, co-immunoprecipitation or pull-down affinity-purification without significant loss in binding capacity.

Applications:
• Immunoprecipitation followed by electrophoresis and excision of the protein for mass spectrometry or sequencing
• Immunoprecipitation and SDS-PAGE analysis of a protein with a molecular weight similar to the antibody's heavy or light chain
• Identification of binding partners of immobilized glycoprotein
• Co-immunoprecipitation for determining protein-protein interactions

Includes:
Kit contains resin, buffers and reagents for immobilization; desalting columns; IP lysis/wash and elution buffers for immunoprecipitation; microcentrifuge spin columns and collection tubes; and loading buffer for SDS-PAGE

The GlycoLink IP Kit method results in covalent attachment of oxidized sugar groups of an IP antibody (or any other glycoprotein with oxidizable carbohydrate groups) to the hydrazide resin. The antibody resin is then incubated with the sample that contains the protein antigen of interest, allowing the antibody-antigen complex to form. After washing to remove non-bound components of the sample, the antigen is recovered by dissociation from the antibody with elution buffer supplied in the kit. The entire procedure is performed in a microcentrifuge spin cup, allowing solutions to be fully separated from the resin with a brief centrifugation. Only the antigen is eluted by the procedure, enabling it to be identified and further analyzed without interference from antibody fragments.

More Product Data
Improved immobilization and conjugation of glycoproteins

96-Well Plates for Pierce™ BCA-RAC Assay (Thermo Scientific™)

These are polystyrene 96-well round-bottom plates suitable for use with Thermo Scientific BCA-RAC (reducing agent compatible) assays.

Related Products
Pierce™ Microplate BCA Protein Assay Kit - Reducing Agent Compatible
Pierce™ BCA Protein Assay Kit - Reducing Agent Compatible

Pierce™ Rapid Antibody Isotyping Kit - Mouse (Thermo Scientific™)

The Thermo Scientific Pierce Rapid Mouse Antibody Isotyping Kit is a single-use, cassette-based kit for 5-minute lateral-flow assay determination of mouse monoclonal antibody class and subclass identity.

This antibody isotyping kit uses cartridges that provide a color-readout of the monoclonal antibody isotype within five minutes after pipetting a small amount of diluted culture supernatant or ascites fluid sample into the sample well. The kit determines mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM.

Features of the Pierce Rapid Mouse Antibody Isotyping Kit:

Long shelf life—stable for at least one year at room temperature
Single-step—add diluted antibody sample to the loading-well of cassette
Fast—within 5 minutes a band appears that indicates the antibody isotype
Sensitive—detect antibodies in tissue culture media or ascites fluid samples at any concentration greater than 10 ng/mL
Specific—assay does not cross-react with fetal bovine serum (FBS)
Reliable—results are similar to the standard ELISA-based isotyping assays

Each Pierce Rapid Isotyping Kit assay is performed by simply adding a properly diluted tissue culture supernatant or mouse ascites sample to the well of the small cassette. Gold conjugates embedded in the cassette form specific class- and subclass-soluble complexes with the antibodies in the sample. These complexes travel the length of the membrane and are resolved on the anti-isotype and class-specific antibody-impregnated membrane. Results are displayed as a red band indicating the antibody isotype or subclass.

Determining the class and subclass of a monoclonal antibody is useful in planning the best immunoglobulin purification method. For example, mouse IgA and IgM are best purified by size (i.e., gel exclusion) or using immunoaffinity separation columns. Mouse IgG2a and IgG2b are purified with immobilized Protein A at pH 7-8, while Mouse IgG1 binds best to Protein A at pH 8-9. Immunoglobulin that contains kappa light chains can be purified using immobilized Protein L.

The Pierce Rapid Isotyping Kit is compatible with both tissue culture supernatant and mouse ascites fluid, and is more sensitive than conventional latex-bead based dipstick assays and much faster than ELISA-based isotyping assays. The kit requires only 0.5µL ascites fluid, 5 to 55µL of cell culture supernatant or 1.5ng of purified antibody.

Related Products
Rapid ELISA Mouse mAb Isotyping Kit
Pierce™ Rapid Antibody Isotyping Kit plus Kappa and Lambda - Mouse

Pro-Detect™ Rapid Antibody Isotyping Assay Kit, rat (Thermo Scientific™)

The Thermo Scientific Pro-Detect Rapid Antibody Isotyping Assay Kit - Rat is a single-use, dipstick lateral flow kit for quick, easy determination of rat monoclonal antibody class and subclass identity. This antibody isotyping assay uses membrane-based strips coated with dye-bound conjugated antibodies that provide a visual, color readout of the monoclonal antibody isotype within ten minutes. The kit determines rat IgG1, IgG2a, IgG2b , and IgG2c.

Features of the Pro-Detect Rapid Antibody Isotyping Assay Kit:
Long shelf life—stable for at least one year at 4°C
Simple workflow—dilute antibody sample using provided diluent and dip in the lateral flow strip
Fast—within 5–10 minutes a band appears that indicates the antibody isotype
Sensitive—detect antibodies in tissue culture media or ascites fluid samples down to low ng/mL concentrations
Specific—assay does not cross-react with fetal bovine serum (FBS)
Reliable—results are similar to standard ELISA-based isotyping assays

Each Pro-Detect Rapid Antibody Isotyping assay is performed by simply adding a properly diluted tissue culture supernatant or mouse ascites sample to a test tube or microplate well and dipping in a lateral flow strip. Gold conjugates embedded in the strip membrane form specific class- and subclass-soluble complexes with the antibodies in the sample. These complexes travel the length of the membrane and are resolved on the anti-isotype and class-specific antibody-impregnated regions of the membrane. Results are displayed as a red band above the printed class or sub-class description, indicating the antibody isotype present.

Determining the subclass of a rat antibody is important when selecting the best purification method. IgG2a, IgG2b, and IgG2c can be purified with Protein A columns at a pH of 7–8. IgG1 isotypes bind best to Protein A at a slightly higher pH of 8–9.

The Pro-Detect Rapid Antibody Isotyping Assay Kit is compatible with both tissue culture supernatant and mouse ascites fluid, and is more sensitive than conventional latex bead-based dipstick assays and much faster than ELISA-based isotyping assays. The kit requires only 0.5 µL ascites fluid, 2–22 µL of cell culture supernatant or 1.5 ng of purified antibody.