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New ABfinity antibodies for ISGylation!

Interferon-stimulated gene 15 (ISG15) encoded by the ISG15 gene is the first identified protein from a family of ubiquitin-like modifiers that share sequence and conformational homology to ubiquitin. The conjugation of ISG15 (Fig a, Fig b) to its target proteins is possible through a three-step process termed as ISGylation which is similar to ubiquitination. In analogy to the ubiquitin system, ISG15 conjugation or ISGylation is mediated by an enzymatic cascade consisting of an E1 activating enzyme UBE1L (Fig c); an E2 conjugating enzyme UBE2L6 and E3 ligases HERC5 (Human) (Fig d)/ HERC6 (Mouse)/ EFP/ HHARI.

Unlike ubiquitination however, ISGylation does not appear to mark proteins for proteasomal degradation; its biological function remains largely undiscovered on account of only a few ISG15 target proteins being identified till now. Considering that type I interferons (IFNs) play critical roles in innate immunity responses and cancer development, ISG15 conjugation is likely to be relatedto IFN mediated immune responses.

 Click images to enlarge

Detection of the free form of Human ISG15 and its conjugates in A549 cells treated with IFN-alpha using ABfinity Anti-ISG15 Human Monoclonal Antibody (Cat. No. 703131, 1:5000) in western blot (Fig a) and (Cat. No. 703654, 1:100) in Immunofluorescence (Fig b).

Detection of UBE1L in A549 and HeLa cells treated with IFN-beta using ABfinity Anti-UBE1L Oligoclonal Antibody (Cat. No. 712170, 1:250) in western blot (Fig c).

Detection of HERC5 in HeLa cells upon siRNA transfection using ABfinity Anti-HERC5 Monoclonal Antibody (Cat. No. 703675, 1:500) in western blot (Fig d).



Click in the table to find out more about or order the ABfinity antibodies for ISGylation

Target Antibody Cat. No
UBE1L 712170
UBE2L6 703136
HERC5 703675
ISG15 (Human) 703131; 703654
ISG15 (Mouse) 703132

Find more antibodies for your research using our antibodies search tool at thermofisher.com/antibodies


Introducing ChIP-grade ABfinity antibodies for mutant histone variants

Histone variants differ from their canonical counterparts in one or a few amino acids that confer specific structural and functional features to these variants. One such histone variant, histone H3.3 is deposited in a replication-independent manner and is associated with transcriptionally active chromatin. Recurrent driver mutations in the histones, predominantly H3.3, trigger genome-wide epigenetic changes, and are common features of certain cancers. These mutations include K27M, G34R/G34V, and K36M. The substitution of glycine at position 34 (G34) modulates methylation at K36, and thus affects transcriptional elongation.

Data from our ChIP-grade monoclonal and oligoclonal recombinant ABfinity H3.3G34R antibodies (Cat. No. 703360, 711969) antibodies are shown below.

Find these and other antibodies for your research using our antibodies search tool at thermofisher.com/antibodies.


New Invitrogen antibodies for flow cytometry

Thermo Fisher Scientific now offers >13,000 Invitrogen antibodies for flow cytometry. With product releases every month, you never know what you might find! Check out our search experience that:

  • Helps you quickly identify the most referenced clone
  • Shows you images directly from the literature
  • Allows you to click directly through to the articles which have used the antibody that is of interest to you

Thermo Fisher Scientific now offers over 685 eBioscience Super Bright antibody conjugates for the violet laser! Additionally, check out other recently released clones:

Use our new tool to search now for your flow cytometry antobodies at thermofisher.com/flowantibodies

Need help finding and selecting the right antibodies for your research? Design your flow cytometry panel now with the Invitrogen Flow Cytometry Panel Builder.


Invitrogen ProQuantum High-Sensitivity Immunoassay Kits - 35 now available

Invitrogen ProQuantum high-sensitivity immunoassays are a platform innovation that provides researchers with an easy-to-run, high-performance assay, with no proprietary instrument to purchase. By utilizing proximity-based amplification technology, we have combined the analyte specificity of high affinity antibody–antigen binding with the signal detection and amplification capabilities of qPCR, to achieve a highly sensitive protein quantitation assay. With the ability to detect even lower levels of protein than traditional methods and with very low sample consumption, you can get the most out of your precious or limited samples.

  • High sensitivity—detect low levels of protein with greater sensitivity than traditional methods
  • Broad dynamic range—≥5 orders of magnitude, minimizing sample dilutions to ensure they fall within the range
  • Low sample consumption—use 2–5 μL of sample (for example, 2 μLvs. 150 μL for triplicate wells with other methods)
  • Fast, easy workflow—2 hours from sample to answer, no wash steps, and a single 1-hour incubation
  • Intuitive cloud-based software—comprehensive data analysis and statistical groupwise comparison
ProQuantum Immunoassays

How ProQuantum Immunoassays work

ProQuantum immunoassays utilize a matched pair of target-specific antibodies, each conjugated to a DNA oligonucleotide. During antibody–analyte binding, the two DNA oligos are brought into close proximity, which then allows ligation of the two strands and subsequent creation of a template strand for amplification.

This immunoassay leverages the sensitivity and large dynamic range of Applied Biosystems TaqMan. qPCR technology. The qPCR data file can be imported into the ProQuantum software (apps.thermofisher.com/apps/proquantum) for easy standard curve analysis and results.

Most popular targets:

Target Cat. No.
Mouse IFNg A41150
Human IFNg A35576
Human IL-5 A35588
Human IL-6 A35573
Human IL-8 A35575
Human TNFa A35601

See the complete list of available protein targets and those coming soon and learn more at thermofisher.com/proquantum.


New AlamarBlue HS and PrestoBlue HS Cell Viability Reagents

The alamarBlue and PrestoBlue HS (high sensitivity) reagents are a new generation of assays that offer superior performance than the first generation of alamarBlue and PrestoBlue reagents. These assays detect cell viability based on cellular redox potential (conversion of resazurin to resofurin) and are compatible with fluorescence- or absorbance-based instruments.

Key advantages:

  • Removal of contaminations from resazurin — displays a >50% reduction in background fluorescence
  • Signal-to-background ratio increased by >100% — resulting in large dynamic range
  • Highly sensitive with a linear response — detects as few as 10 cells per well
  • Convenient add-and-read format — no mixing, no washing, no cell lysis, and compatible with either fluorescence- or absorbance-based instrumentation

Comparison between alamarBlue HS and PrestoBlue HS

  alamarBlue HS PrestoBlue HS
Contains highly purified resazurin, resulting in low background and higher signal-to-background ratio Yes Yes
Works with diverse cells types (mammalian, fungal, plant, bacterial, etc.) Yes  
Optimized for mammalian cells   Yes
Longer experiments or higher cell densities Yes  
Faster determination of viability (~10 minutes)   Yes
PrestoBlue HS Cell Viability Reagent P50201
alamarBlue HS vs competition

Performance comparison of alamarBlue cell viability reagents. A549 (left panel) or HeLa (right panel) cells were seeded at 5,000 cell/well and incubated overnight to allow for attachment. Only complete media was added to the Background wells. A 10 µL aliquot of alamarBlue HS (Thermo Fisher Scientific), alamarBlue (Thermo Fisher Scientific) or alamarBlue (Bio-Rad) reagent was added to the wells containing 100 µL of complete growth media and the attached cells. At various time points post-addition of the alamarBlue cell health reagents the fluorescence was measured using at excitation 560nm and an emission of 590nm. The Signal to Background ratio was generated by subtracting the Background fluorescence (media and alamarBlue cell health reagent) from the Signal fluorescence (cells, media and alamarBlue cell health reagent). Within 30 minutes the increase in the Signal to Background ratio for the alamarBlue HS is clearly visible and increases over time for both cell lines. After four hours of incubation the A549 cells treated with alamarBlue HS displays a Signal to Background ratio of >200-fold; when compared to the ratio for alamarBlue (from Thermo Fisher or Bio-Rad) of 20-fold this is a >100% increase in the signal to background ratio for the alamarBlue HS versus the alamarBlue reagent.

If you are currently using alamarBlue and PrestoBlue reagents (DAL1025, DAL1100, A13261, A13262), you can try these new HS versions.

Product Name Cat. No. Unit size
alamarBlue HS Cell Viability Reagent A50100 25 mL
A50101 100 mL
PrestoBlue HS Cell Viability Reagent P50200 25 mL
P50201 100 mL

Learn more about cell viability assays


CellEvent Senescence Kits for imaging and flow cytometry

The CellEvent Senescence Reagent kits are optimized for the fluorescent detection of senescent cells. Upregulation and accumulation of the β-galactosidase enzyme (β-gal) is commonly used as a biomarker for senescent cells. This hydrolase enzyme resides in lysosomes and converts β-galactosides into monosaccharides under acidic pH conditions.

The kits' fluorescent probe contains two galactoside moieties, making it specific for β-galactosidase. The enzymatically cleaved product is retained within the cell due to covalent binding of intracellular proteins and emits a fluorogenic signal that has absorption/emission maxima of 490/514 nm.

These kits offer:

  • Reliable and quick fluorescence detection of senescent cells
  • Simply add, incubate, and detect using standard Alexa Fluor 488/FITC filter sets
  • Multiplexing – fluorescent senescence probe can be multiplexed
  Product Name Cat. No. Unit size
Imaging CellEvent Senescence Green Detection Kits C10850 25 µL
C10851 100 µL
Flow cytometry CellEvent Senescence Green Flow Cytometry Assay Kits C10840 50 assays
C10841 200 assays
AlamarBlue HS Cell Viability Reagent P50101

New cloud library protocols for antibody workflows in microplate readers

Antibody-related applications are now even more straightforward with the aid of the Cloud Library in the Thermo Scientific SkanIt Software 6.0, which controls all Thermo Scientific microplate readers. The SkanIt Cloud Library is a collection of validated sessions, with real assay data and calculations, that all users can access for performing immediate measurements, or as templates to create their own assay protocols. With SkanIt Software 6.0, the Cloud Library is now available in a redesigned format, making it simpler to search for protocols by instrument, detection technology, plate format or other relevant search terms. The library is also expanded to cover more than 130 sessions. Each Cloud Library session has complete measurements with the required calculation and graph processing steps as well as representative data from real assays. Instructions for running the assays are also provided.

Ready-made cloud sessions are available for quick estimations of concentrations of antibodies using absorbance measurements at 280 nm. Assays can be selected to be performed with the Thermo Scientific μDrop Plate (for micro volumes) or with 96-well plates, using Thermo Scientific Multiskan Sky Spectrophotometer or Varioskan LUX Multimode Microplate Reader. When characterizing dye-conjugated antibodies, fast fluorometric titrations can be carried out with the dedicated protocols of Thermo Scientific Varioskan LUX, Fluoroskan or Fluoroskan FL microplate readers. Spectral measurements are also included with Varioskan LUX, for antibody validation purposes.

Learn more about SkanIt 6.0 Software at thermofisher.com/skanit

To compare Thermo Scientific microplate readers driven by SkanIt Software, visit thermofisher.com/platereaders

NEWS AND VIEWS

Alexa Fluor Plus secondary antibodies- new testing data

Visit the updated web page for these secondary antibody conjugates launched at the end of 2018. These innovative secondary antibody conjugates combine the performance of Alexa Fluor Plus dyes with the cross-adsorbed secondary antibodies to deliver greater sensitivity and best signal-to background ratio. Review the testing data or search for the antibodies you need for your application and/or multiplexing.

Comparison product data is now available, along with an easy to use Alexa Fluor Plus secondary antibody ordering table.

  Invitrogen Alexa Fluor Plus Secondary Antibody products
Click on the catalog number below to choose.
Goat anti-Rabbit Goat anti-Mouse Goat anti-Chicken Donkey anti-Goat Donkey anti-Rabbit Donkey anti-Mouse
Alexa Fluor Plus 488 A32731 A32723 A32931 A32814 A32790 A32766
Alexa Fluor Plus 555 A32732 A32727 A32932 A32816 A32794 A32773
Alexa Fluor Plus 594 A32740 A32742 A32759 A32758 A32754 A32744
Alexa Fluor Plus 647 A32733 A32728 A32933 A32849 A32795 A32787
Alexa Fluor Plus 680 A32734 A32729 A32934 A32860 A32802 A32788
Alexa Fluor Plus 800 A32735 A32730 A32935 A32930 A32808 A32789

Read the blog: Macrophages by flow cytometry

Do you have questions about macrophages? What are they? What do they do? Read the blog post where we talk with our expert, Dariusz Stepniak, Ph.D., a Staff Scientist in R&D at Thermo Fisher Scientific to answer these questions. Additional topics covered include a discussion about macrophage classification, the difference between M1 and M2 macrophages and key markers to consider when characterizing macrophages. A few key references about macrophages are also provided.

Read the blog post about macrophages


Read the blog: New insights through the RELM alpha antibody

The Invitrogen anti-mouse RELM alpha monoclonal antibody (clone DS8RELM) recognizes RELM alpha, also known as resistin-like alpha or FIZZ1 a small cytokine produced mostly by macrophages. This antibody is particularly valuable to the scientists who aim to better understand the process of macrophage polarization. For more information about the DS8RELM clone, please read the interview with Dariusz Stepniak, Ph.D., a Staff Scientist in R&D at Thermo Fisher Scientific, who was responsible for the development of this new antibody.

Dariusz Stepniak

The new Invitrogen ELISA web experience

The new Invitrogen ELISA web experience

Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.