PE anti-human CD279 (PD-1) Antibody

Product Details

Verified Reactivity

Human

Reported Reactivity

African Green, Baboon, Chimpanzee, Common Marmoset, Cynomolgus, Rhesus, Squirrel Monkey

Antibody Type

Monoclonal

Host Species

Mouse

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)

Preparation

The antibody was purified by affinity chromatography, and conjugated with PE under optimal conditions.

Concentration

Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)

Storage & Handling

The CD279 antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.

Application

FC - Quality tested
SB - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

Excitation Laser

Blue Laser (488 nm)
Green Laser (532 nm)/Yellow-Green Laser (561 nm)

Application Notes

Additional reported applications (for the relevant formats) include: blocking of ligand binding^1-3, immunohistochemical staining of paraformaldehyde fixed frozen sections¹³, and spatial biology (IBEX)^15,16. The LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 329911 and 329912). For highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 329926) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin <0.01 EU/µg).

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References

1. Dorfman DM, et al. 2006 Am. J. Surg. Pathol. 30:802. (FA)
2. Radziewicz H, et al. 2007. J. Virol. 81:2545. (FA)
3. Velu V, et al. 2007. J. Virol. 81:5819. (FA)
4. Zahn RC, et al. 2008. J. Virol. 82:11577. PubMed
5. Chang WS, et al. 2008. J. Immunol. 181:6707. (FC) PubMed
6. Nakamoto N, et al. 2009. PLoS Pathog. 5:e1000313. (FA)
7. Jones RB, et al. 2009. J. Virol. 83:8722. (FC) PubMed
8. Vojnov L, et al. 2010. J. Virol. 84:753. (FC) PubMed
9. Radziewicz H, et al. 2010. J. Immunol. 184:2410. (FC) PubMed
10. Monteriro P, et al. 2011. J. Immunol. 186:4618. PubMed
11. Conrad J, et al. 2011. J. Immunol. 186:6871. PubMed
12. Salisch NC, et al. 2010. J. Immunol. 184:476. (Rhesus reactivity)
13. Li H and Pauza CD. 2015. Eur. J. Immunol. 45:298. (IHC)
14. Peterson VM, et al. 2017. Nat. Biotechnol. 35:936. (PG)
15. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
16. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed

Product Citations

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RRID

AB_940481 (BioLegend Cat. No. 329905)
AB_940481 (BioLegend Cat. No. 329906)

Antigen Details

Structure

Immunoglobulin superfamily

Distribution

Transiently expressed on CD4^- CD8^- thymocytes; upregulated in thymocytes and splenic T and B lymphocytes; expressed on activated myeloid cells

Ligand/Receptor

B7-H1 (also known as PD-L1) and B7-DC (PD-L2)

Cell Type

B cells, Lymphocytes, T cells, Thymocytes, Tregs

Biology Area

Cancer Biomarkers, Immunology, Inhibitory Molecules

Molecular Family

CD Molecules, Immune Checkpoint Receptors

Gene ID

5133 View all products for this Gene ID

Specificity (DOES NOT SHOW ON TDS):

CD279

Specificity Alt (DOES NOT SHOW ON TDS):

CD279

App Abbreviation (DOES NOT SHOW ON TDS):

FC,SB

UniProt

View information about CD279 on UniProt.org

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Material Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

Related FAQs

What type of PE do you use in your conjugates?

We use R-PE in our conjugates.

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Other Formats

Certificate of Analysis