Alexa Fluor® 647 anti-mouse CD4 Antibody

Pricing & Availability
Clone
GK1.5 (See other available formats)
Other Names
L3T4, T4
Isotype
Rat IgG2b, κ
Ave. Rating
2 reviews
Product Citations
publications
A_GK1.5_AF647_021706
C57BL/6 mouse splenocytes were stained with CD4 (clone GK1.5) Alexa Fluor® 647 (filled histogram) or rat IgG2b, κ Alexa Fluor® 647 isotype control (open histogram).
  • A_GK1.5_AF647_021706
    C57BL/6 mouse splenocytes were stained with CD4 (clone GK1.5) Alexa Fluor® 647 (filled histogram) or rat IgG2b, κ Alexa Fluor® 647 isotype control (open histogram).
  • B_GK1.5_A647_CD4_IHCF_040318
    C57BL/6 mouse frozen lymph node section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS plus 5% rat serum for 1 hour at room temperature. Then the section was stained with 5 µg/ml of CD8 (clone 53-6.7) Alexa Fluor® 594 (red), 5 µg/ml of CD4 (clone GK1.5) Alexa Fluor® 647 (blue), and 5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (green) overnight at 4°C. The image was captured by 10X objective.
See Alexa Fluor® 647 spectral data
Cat # Size Price Quantity Avail. Save
100426 25 µg $85
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100424 100 µg $195
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Description

CD4 is a 55 kD protein also known as L3T4 or T4. It is a member of the Ig superfamily, primarily expressed on most thymocytes, a subset of T cells, and weakly on macrophages and dendritic cells. It acts as a coreceptor with the TCR during T cell activation and thymic differentiation by binding MHC class II and associating with the protein tyrosin kinase, lck.

Product Details
Technical data sheet

Product Details

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 Alexa Fluor® 647 under optimal conditions. The solution is free of unconjugated Alexa Fluor® 647.
Concentration
0.5 mg/ml
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
IHC-F - Validated

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 million cells in 100 µl volume. For immunohistochemistry of frozen tissue sections, a concentration range of 2.5-5 μg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Application Notes

Additional reported applications (for the relevant formats) include: blocking of CD4+ T cell activation1,4,11, thymocyte costimulation3, in vitro and in vivo depletion2,5-8, blocking of egg-sperm cell adhesion1,4, immunohistochemical staining of acetone-fixed frozen sections9,10, and immunoprecipitation1,2. 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 (Cat. No. 100506), but not RM4-4 antibody (Cat. No. 116002). The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm filtered) is recommended for functional assays (Cat. No. 100416). 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).

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)
Product Citations
  1. Li J, et al. 2012. Arthritis Rheum. 64:1098. PubMed
  2. Ishida W, et al. 2014. Clin Immunol. 153:136. PubMed
  3. Sebastian M, et al. 2016. J Immunol. 196: 144 - 155. PubMed
  4. Cooley L, et al. 2016. Sci Rep. 6: 25840. PubMed
  5. Mier-Aguilar C, et al. 2016. PLoS One. 11:e0168155. PubMed
  6. Lin JR 2018. eLife. 7: e31657. PubMed
  7. Uchil PD, et al. 2019. Cell Host Microbe. 25:87. PubMed
  8. Zhang YS, et al. 2018. Cancer Biol Ther. 19:735. PubMed
  9. Palmer BC, et al. 2019. Sci Rep. 9:5085. PubMed
  10. Wang F, et al. 2018. Carcinogenesis. 39:889. PubMed
Publication Library
RRID
AB_493519 (BioLegend Cat. No. 100426)
AB_389324 (BioLegend Cat. No. 100424)

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

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.
Go To Top Version: 3    Revision Date: 06/27/2014

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*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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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.
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