The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Application:
FC
Recommended Usage:
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.
* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome. ** Pacific Blue™ is a registered trademark of Molecular Probes, Inc. Pacific Blue™ dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
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 Pacific Blue™ GK1.5
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.
