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.
Storage & Handling:
The antibody solution should be stored undiluted at 4 °C. This LEAF™ solution contains no preservative; handle under aseptic conditions.
Application:
FC - Quality Tested Blocking1,2,4*, IHC(Frozen)3*, IP4, WB(non-reducing)* - *This application has been reported in the literature.
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 ≤ 0.5 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: in vitro and in vivo blocking of cell-cell adhesion and CD54 functions1,2,4, immunohistochemical staining3 of acetone-fixed frozen sections, immunoprecipitation4, and Western blotting (non-reducing). The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 116110).
Application References:
1. Kumaska, T., et al., 1996. J. Clin. Invest. 97:2362. 2. Horley, K.J., et al., 1989. EMBO J. 8:2889. 3. Burns, A.R., et al., 1994. J. Immunol. 153:3189. 4. Takei, F., et al., 1985. J. Immunol. 134:1403. 5. Nishimura., et al. 2008. J. Clin Invest. 118:710. PubMed
C57BL/6 mouse splenocytes stained with LEAF™ purified YN1/1.7.4 , followed by anti-rat IgG FITC
Description:
CD54 is a 90 kD immunoglobulin superfamily member, also known as ICAM-1 and Ly-47. It is expressed on activated endothelial cells, high endothelial venules (HEV), T and B cells, monocytes/ macrophages, granulocytes, and dendritic cells. CD54 is an important intracellular adhesion molecule that participates in T cell-T cell, T cell-B cell, and T cell-target cell interactions via binding of LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18). CD54 has also been shown to be involved in lymphocyte trafficking, making it an important molecule in many immune reactions and inflammation. CD54 is also a receptor for rhinovirus. The YN1/1.7.4 antibody has been reported to block binding of mouse CD54 to LFA-1 and Mac-1, and inhibits cell-cell adhesion, antigen presentation to T cells, and leukocyte migration to inflammatory tissues.
Other Names:
ICAM-1, Ly-47
Structure:
Ig superfamily, 90 kD
Distribution:
Activated endothelial cells, high endothelial venules, T and B cells, monocytes/macrophages, granulocytes, dendritic cells
Function:
Immune reaction, inflammation, adhesion
Ligand Receptor:
CD11a/CD18 (LFA-1) or CD11b/CD18 (Mac-1) and CD11c/CD18, CD43, hyaluronan, fibrinogen
Antigen References:
1. Barclay, A.N., et al., 1997. The Leukocyte Antigen FactsBook, Academic Press. 2. Springer, T.A. 1990. Nature 346:425. 3. Rothlein, R., et al., 1986. J. Immunol. 137:1270.
