Brilliant Violet 421™ anti-mouse CD279 (PD-1) Antibody

Pricing & Availability
Clone
29F.1A12 (See other available formats)
Regulatory Status
RUO
Other Names
PD-1, Programmed Death-1
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
29F.1A12_BV421_1_062911
Con-A and IL-2 stimulated C57BL/6 splenocytes (3 days) were stained with CD3 APC and CD279 (clone 29F.1A12) Brilliant Violet 421™ (top), or rat IgG2a, κ Brilliant Violet 421™ isotype control (bottom).
  • 29F.1A12_BV421_1_062911
    Con-A and IL-2 stimulated C57BL/6 splenocytes (3 days) were stained with CD3 APC and CD279 (clone 29F.1A12) Brilliant Violet 421™ (top), or rat IgG2a, κ Brilliant Violet 421™ isotype control (bottom).
  • 29F.1A12_BV421_2_062911
  • 60_Mouse_Lymph_Node_PD1_CD23_IgD
    Mice were injected subcutaneously with sheep red blood cells in a volume of 25 µl per site on days 0 and 4 and harvested on day 11. Confocal image of C57BL/6 mouse lymph node acquired using the IBEX method of highly multiplexed antibody-based imaging: PD-1 (green) in Cycle 1, CD23 (magenta) in Cycle 7, and IgD (blue) in Cycle 10. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Compare all formats See Brilliant Violet 421™ spectral data
Cat # Size Price Quantity Check Availability Save
135217 125 µL 164€
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135221 50 µg 208€
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135218 500 µL 322€
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Description

CD279, also known as programmed death-1 (PD-1), is a 50-55 kD glycoprotein belonging to the CD28 family of the Ig superfamily. PD-1 is expressed on activated splenic T and B cells and thymocytes. It is induced on activated myeloid cells as well. PD-1 is involved in lymphocyte clonal selection and peripheral tolerance through binding its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2). It has been reported that PD-1 and PD-L1 interactions are critical to positive selection and play a role in shaping the T cell repertoire. PD-L1 negative costimulation is essential for prolonged survival of intratesticular islet allografts.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
PD-1 cDNA followed by PD-1-Ig fusion protein
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 Brilliant Violet 421™ under optimal conditions.
Concentration
µg sizes: 0.2 mg/mL
µL sizes: lot-specific (to obtain lot-specific concentration, please enter the lot number in our Concentration and Expiration Lookup or Certificate of Analysis online tools.)
Storage & Handling
The 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 immunofluorescent staining using the µg size, the suggested use of this reagent is ≤0.125 µg per million cells in 100 µl volume. For immunofluorescent staining using µl sizes, 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. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.


Learn more about Brilliant Violet™.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.
Excitation Laser
Violet Laser (405 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen tissue3, in vivo blocking of PD-1 binding to its ligands2,3, and spatial biology (IBEX)5,6.

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

(PubMed link indicates BioLegend citation)
  1. Good-Jacobson KL, et al. 2010. Nat. Immunol. 11:535. (FC) PubMed
  2. Lázár-Molnár E, et al. 2008. Proc. Natl. Acad. Sci. USA 105:2658. (Block)
  3. Liang SC, et al. 2003. Eur. J. Immunol. 33:2706. (FC, IHC, Block)
  4. Tobias J, et al. 2020. Front Immunol. 11:895 (FC, ELISA) PubMed
  5. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  6. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Martínez‐López M et al. 2019. Immunity. 50(2):446-461 . PubMed
  2. Nagai Y, et al. 2019. Front Immunol. 10:174. PubMed
  3. Tavazoie MF, et al. 2018. Cell. 172:825. PubMed
  4. Wei SC, et al. 2019. Immunity. 50:1084. PubMed
  5. Papa I, et al. 2017. Nature. 547:318. PubMed
  6. Pein M, et al. 2020. Nat Commun. 11:1494. PubMed
  7. Liu H, et al. 2020. Cancer Cell. 37(3):324-339. PubMed
  8. Kinsey G, et al. 2012. J Am Soc Nephrol. 23:1528. PubMed
  9. Kim Y, et al. 2015. PLoS One. 10:120294. PubMed
  10. Wan X, Thomas J, Unanue E 2016. J Exp Med. 213: 967 - 978. PubMed
  11. Scala M, et al. 2016. J Virol. 90: 8563 - 8574. PubMed
  12. RY H, et al. 2016. Oncoimmunology. 6:e1249561. PubMed
  13. Ye Y, et al. 2020. Genome Med. 0.557638889. PubMed
  14. Clemmensen HS, et al. 2020. Front Immunol. 11:585359. PubMed
  15. Calvo-Barreiro L, et al. 2021. Neurotherapeutics. . PubMed
  16. Nicolas-Boluda A, et al. 2021. eLife. 10:00. PubMed
  17. Kumagai S, et al. 2020. Immunity. 53(1):187-203.e8. PubMed
  18. Gonzalez-Figueroa P, et al. 2021. Cell. 184(7):1775-1789.e19. PubMed
  19. Delacher M, et al. 2021. Immunity. 54(4):702-720.e17. PubMed
  20. Wong HS, et al. 2021. Cell. . PubMed
RRID
AB_10900085 (BioLegend Cat. No. 135217)
AB_2562568 (BioLegend Cat. No. 135221)
AB_2561447 (BioLegend Cat. No. 135218)

Antigen Details

Structure
A 50-55 kD glycoprotein belonging to the CD28 family of the Ig superfamily.
Distribution

Induced on splenic T and B lymphocytes, thymocytes, and myeloid cells after stimulation.

Function
Involved in lymphocyte clonal selection and peripheral tolerance, prolonged survival of allografts.
Ligand/Receptor
B7-H1 (PD-L1) and B7-DC (PD-L2)
Cell Type
B cells, T cells
Biology Area
Cancer Biomarkers, Immunology, Inhibitory Molecules
Molecular Family
CD Molecules, Immune Checkpoint Receptors
Antigen References

1. Nishimura H, et al. 2001. Science 291:319
2. Agata Y, et al. 1996. Int. Immunol. 8:765
3. Liang SC, et al. 2003. Eur. J. Immunol. 33:2706
4. Barber DL, et al. 2006. Nature 439:682
5. Keir ME, et al. 2005. J. Immunol. 175:7372
6. Koehn BH. et al. 2008. J Immunol. 181:5313

Gene ID
18566 View all products for this Gene ID
UniProt
View information about CD279 on UniProt.org

Related FAQs

What is the F/P ratio range of our BV421™ format antibody reagents?

It is lot-specific. On average it ranges between 2-4.

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.

Go To Top Version: 4    Revision Date: 04/19/2022

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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