PerCP/Cyanine5.5 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.

Preparation

The antibody was purified by affinity chromatography, and conjugated with PerCP/Cyanine5.5 under optimal conditions.

Concentration

0.2 mg/ml

Storage & Handling

The CD4 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 flow cytometric staining, the suggested use of this reagent is = 0.25 µg per 10⁶ cells in 100 µl. It is recommended that the reagent be titrated for optimal performance for each application.

* PerCP/Cyanine5.5 has a maximum absorption of 482 nm and a maximum emission of 690 nm.

Excitation Laser

Blue Laser (488 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

BioLegend is in the process of converting the name PerCP/Cy5.5 to PerCP/Cyanine5.5. The dye molecule remains the same, so you should expect the same quality and performance from our PerCP/Cyanine5.5 products. Contact Technical Service if you have any questions.

Application References

(PubMed link indicates BioLegend citation)

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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

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RRID

AB_893330 (BioLegend Cat. No. 100433)
AB_893324 (BioLegend Cat. No. 100434)

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

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.

How stable is PerCP/Cyanine5.5 tandem as compared to PerCP alone?

PerCP/Cyanine5.5 is quite photostable and also better than PerCP alone in withstanding fixation.

Other Formats

Certificate of Analysis