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:
This reagent is developed for immunofluorescent staining for flow cytometric analysis, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. It is highly 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: immunoprecipitation4, in vitro blocking of B cell responses1,2, and immunohistochemical staining3 of acetone-fixed frozen sections. The A20 antibody does not react with mouse cells expressing the CD45.2 alloantigen.
Application References:
1. Yakura, H., et al., 1983. J. Exp. Med. 157:1077. 2. Yakura, H., et al., 1986. J. Immunol. 136:2729. 3. Suzuki, K., et al., 2000. Immunity 13:691. 4. Shen, F.-W., et al., 1986. Immunogenetics 24:146. 5. Pascal, V., et al. 2007. J. Immunol. 179:1751. 6. Phan, T.G., et al., 2007. Nature Immunol. 8:992. 7.Beamer,CA.,et al.2007. Am J Respir Cell Mol Biol. 37:729.PubMed
SJL mouse splenocytes stained with A20 Pacific Blue™
Description:
CD45.1 is an alloantigen of CD45, expressed by Ly5.1 bearing mouse strains (e.g., RIII, SJL/J, STS/A, DA). CD45, a member of the protein tyrosine phosphatase (PTP) family, is a 180-240 kD glycoprotein expressed on all hematopoietic cells except mature erythrocytes and platelets. There are multiple isoforms in the mouse that play key roles in TCR and BCR signal transduction. These isoforms are very specific to the activation and maturation states of the cell, as well as specific cell type. The primary ligands for CD45 are galectin-1, CD2, CD3, CD4, TCR, CD22, and Thy-1. The A20 antibody does not react with leukocytes that express the CD45.2 alloantigen.
Other Names:
T200, Ly-5.1, LCA
Structure:
Protein tyrosine phosphatase (PTP) family, 180-240 kD
Distribution:
All hematopoietic cells except mature erythrocytes and platelets of the CD45.1 strain of mice
Function:
Phosphatase, T and B cell activation
Ligand Receptor:
Galectin-1, CD2, CD3, CD4
Antigen References:
1. Barclay, A., et al., 1997. The Leukocyte Antigen FactsBook, Academic Press. 2. Trowbridge, I.S., et al., 1993. Annu. Rev. Immunol. 12:85. 3. Kishihara, K., et al., 1993. Cell 74:143. 4. Pulido, R., et al., 1988. J. Immunol. 140:3851.
