Brilliant Violet 510™ anti-mouse CD4 Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

BALB/c mouse thymocytes

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 510™ 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

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 ≤0.5 µg per million cells in 100 µl volume. For immunofluorescent 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 510™ excites at 405 nm and emits at 510 nm. The bandpass filter 510/50 nm is recommended for detection, although filter optimization may be required depending on other fluorophores used. Be sure to verify that your cytometer configuration and software setup are appropriate for detecting this channel. Refer to your instrument manual or manufacturer for support. Brilliant Violet 510™ 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

The RM4-5 antibody blocks the binding of GK1.5 antibody and H129.19 antibody to CD4⁺ T cells, but not RM4-4 antibody. Additional reported applications (for the relevant formats) include: blocking of ligand binding, in vivo depletion of CD4⁺ cells¹, and immunohistochemistry of acetone-fixed frozen tissue sections^2,3,11 and paraffin-embedded sections¹¹. Clone RM4-5 is not recommended for immunohistochemistry of formalin-fixed paraffin sections. Instead, acetone frozen or zinc-fixed paraffin sections are recommended. The Ultra-LEAF™ Purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 100575 and 100576).

Application References

(PubMed link indicates BioLegend citation)

1. Kruisbeek AM. 1991. In Curr. Protocols Immunol. pp. 4.1.1-4.1.5. (Block, Deplete)
2. Nitta H, et al. 1997. Cell Vision 4:73. (IHC)
3. Fan WY, et al. 2001. Exp. Biol. Med. 226:1045.
4. Muraille E, et al. 2003. Infect. Immun. 71:2704. (IHC)
5. León-Ponte M, et al. 2007. Blood 109:3139. (FC)
6. Bourdeau A, et al. 2007. Blood doi:10.1182/blood-2006-08-044370. (FC)
7. Matsumoto M, et al. 2007.J. Immunol.178:2499. PubMed
8. Shigeta A, et al. 2008. Blood 112:4915. PubMed
9. Zaborsky N, et al. 2010. J. Immunol. 184:725. PubMed
10. Rodrigues-Manzanet R, et al. 2010. P. Natl Acad Sci USA 107:8706. PubMed
11. Whiteland JL, et al. 1995. J. Histochem. Cytochem. 43:313. (IHC)

Product Citations

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RRID

AB_2561388 (BioLegend Cat. No. 100553)
AB_2562608 (BioLegend Cat. No. 100559)

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 (pdf)
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry 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.

Other Formats

Certificate of Analysis