Phosphate-buffered solution, pH 7.2, containing no preservative. 0.2 μm filter sterilized. Endotoxin level is < 0.1 EU/μg of the protein (< 0.01 ng/μg of the protein) as determined by the LAL test.
Preparation:
The LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration:
1 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4 °C. This LEAF™ solution contains no preservative; handle under aseptic conditions.
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
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-cell to sperm cell adhesion1,4, immunohistochemical staining9,10 of acetone-fixed frozen sections, and immunoprecipitation1,2. 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 sterile-filtered) is recommended for functional assays (Cat. No. 100416).
Application References:
1. Dialynas, D.P., et al., 1983. J. Immunol. 131:2445. 2. Dialynas, D.P., et al., 1983. Immunol. Rev. 74:29. 3. Wu, L., et al., 1991. J. Exp. Med. 174:1617. 4. Godfrey, D.I., et al., 1994. J. Immunol. 152:4783. 5. Gavett, S.H., et al., 1994. Am. J. Respir. Cell. Mol. Biol. 10:587. 6. Schuyler, M., et al., 1994. Am. J. Respir. Crit. Care Med. 149:1286. 7. Ghobrial, R.R., et al., 1989. Clin. Immunol. Immunopathol. 52:486. 8. Israelski, D.M., et al., 1989. J. Immunol. 142:954. 9. Zheng, B., et al., 1996. J. Exp. Med. 184:1083. 10. Frei, K., et al., 1997. J. Exp. Med. 185:2177. 11. Felix, N.J., et al., 2007. Nature Immunol. 8:388.
C57BL/6 mouse splenocytes stained with LEAF™ purified GK1.5, followed by anti-rat IgG FITC
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. The GK1.5 antibody is able to block CD4 mediated cell adhesion and T cell activation. It is also useful for in vivo and in vitro depletion of CD4+ T cells.
Other Names:
L3T4, T4
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
Antigen References:
1. Barclay, A., et al., 1997. The Leukocyte Antigen FactsBook, Academic Press. 2. Bierer, B.E., et al., 1989. Annu. Rev. Immunol. 7:579. 3. Janeway, C.A. 1992. Annu. Rev. Immunol. 10:645.
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