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 ligand binding, in vivo depletion of CD4+ cells1, immunohistochemistry2,3 of acetone-fixed frozen sections, and 10% formalin-fixed paraffin-embedded tissue sections3. Special Note: The antibody works well on acetone-fixed frozen sections. It sometime works on formalin-fixed paraffin-embedded tissue sections but inconsistent results have been reported. The RM4-5 antibody blocks the binding of GK1.5 antibody (Cat. No. 100402) to CD4 T cells, but not RM4-4 antibody (116002). The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 100520).
Application References:
1. Kruisbeek, A.M. 1991. In Curr. Protocols Immunol. pp. 4.1.1-4.1.5. 2. Nitta, H.,et al., 1997. Cell Vision 4:73. 3. Fan, W. Y., et al., 2001. Exp. Biol. Med. 226: 1045. 4. Muraille, E., et al., 2003. Infect. Immun. 71:2704. 5. León-Ponte, M., et al., 2007. Blood 109:3139. 6. Bourdeau, A., et al., 2007. Blood doi:10.1182/blood-2006-08-044370.
C57BL/6 mouse splenocytes stained with LEAF™ purified RM4-5, 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 RM4-5 antibody is useful for in vitro blocking of ligand binding, as well as in vivo CD4+ T cell depletions.
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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