Brilliant Violet 421™ anti-mouse CD4 Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

Mouse CTL clone V4

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 and expiration, please enter the lot number in our Certificate of Analysis online tool.)

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
ICC - Verified

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 using the µg size, the suggested use of this reagent is ≤0.125 µg per million cells in 100 µl volume. For flow cytometric staining using the µl size, 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: blocking of CD4⁺ T cell activation^1,4,11, thymocyte costimulation³, in vitro and in vivo depletion^2,5-8, blocking of egg-sperm cell adhesion^1,4, immunohistochemical staining of acetone-fixed frozen sections^9,10, immunoprecipitation^1,2, and spatial biology (IBEX)^12,13. The GK1.5 antibody is able to block CD4 mediated cell adhesion and T cell activation. Binding of GK1.5 antibody to CD4 T cells can be blocked by RM4-5 antibody, but not RM4-4 antibody. For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 100442) 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

(PubMed link indicates BioLegend citation)

1. Dialynas DP, et al. 1983. J. Immunol. 131:2445. (Block, IP)
2. Dialynas DP, et al. 1983. Immunol. Rev. 74:29. (IP, Deplete)
3. Wu L, et al. 1991. J. Exp. Med. 174:1617. (Costim)
4. Godfrey DI, et al. 1994. J. Immunol. 152:4783. (Block)
5. Gavett SH, et al. 1994. Am. J. Respir. Cell. Mol. Biol. 10:587. (Deplete)
6. Schuyler M, et al. 1994. Am. J. Respir. Crit. Care Med. 149:1286. (Deplete)
7. Ghobrial RR, et al. 1989. Clin. Immunol. Immunopathol. 52:486. (Deplete)
8. Israelski DM, et al. 1989. J. Immunol. 142:954. (Deplete)
9. Zheng B, et al. 1996. J. Exp. Med. 184:1083. (IHC)
10. Frei K, et al. 1997. J. Exp. Med. 185:2177. (IHC)
11. Felix NJ, et al. 2007. Nat. Immunol. 8:388. (Block)
12. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
13. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed

Product Citations

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RRID

AB_10900241 (BioLegend Cat. No. 100437)
AB_10900241 (BioLegend Cat. No. 100443)
AB_10900241 (BioLegend Cat. No. 100438)

Antigen Details

Structure

Ig superfamily, 55 kD

Distribution

Majority of thymocytes, T cell subset

Function

TCR co-receptor, T cell activation

Ligand/Receptor

MHC class II molecule

Cell Type

Dendritic cells, T cells, Thymocytes, Tregs

Biology Area

Immunology

Molecular Family

CD Molecules

Antigen References

1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press.
2. Bierer BE, et al. 1989. Annu. Rev. Immunol. 7:579.
3. Janeway CA. 1992. Annu. Rev. Immunol. 10:645.

Gene ID

12504 View all products for this Gene ID

UniProt

View information about CD4 on UniProt.org

Documentation

• Technical data sheet
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol
• Immunocytochemistry Staining Protocol

Related Pages & Pathways

Related FAQs

I am unable to see expression of T cell markers such as CD3 and CD4 post activation.

TCR-CD3 complexes on the T-lymphocyte surface are rapidly downregulated upon activation with peptide-MHC complex, superantigen or cross-linking with anti-TCR or anti-CD3 antibodies. PMA/Ionomycin treatment has been shown to downregulate surface CD4 expression. Receptor downregulation is a common biological phenomenon and so make sure that your stimulation treatment is not causing it in your sample type.

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

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

Other Formats

Certificate of Analysis