BALB/C mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% rat serum + FBS for one hour at room temperature. Then the section was stained with 10 µg/ml of CD45.2 (clone 104) Alexa Fluor® 594 (red), 5 µg/ml of CD3 (clone 17A2) Alexa Fluor® 647 (green), and 5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (blue) at room temperature for two hours. The image was captured with a 10X objective.
BALB/C mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% rat serum + FBS for one hour at room temperature. Then the section was stained with 10 µg/ml of CD45.2 (clone 104) Alexa Fluor® 594 (red), 5 µg/ml of CD3 (clone 17A2) Alexa Fluor® 647 (green), and 5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (blue) at room temperature for two hours. The image was captured with a 10X objective.
CD45.2 is an alloantigen of CD45, expressed by Ly5.2 bearing mouse strains (e.g., A, AKR, BALB/c, CBA/Ca, CBA/J, C3H/He, C57BL, C57BR, C57L, C58, DBA/1, DBA/2, NZB, SWR, 129). 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.
Each lot of this antibody is quality control tested by immunohistochemical staining on frozen tissue sections. For immunohistochemistry, a concentration range of 5.0 - 10 μg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
The 104 antibody does not react with mouse cells expressing the CD45.1 alloantigen. Additional reported applications (for the relevant formats) include: immunoprecipitation4, in vivo and in vitro blocking of B cell responses1,2, and immunohistochemical staining of acetone-fixed frozen sections3.
Application References
(PubMed link indicates BioLegend citation)
Yakura H, et al. 1983. J. Exp. Med. 157:1077. (Block)
Yakura H, et al. 1986. J. Immunol. 136:2729. (Block)
Suzuki K, et al. 2000. Immunity 13:691. (IHC)
Shen FW, et al. 1986. Immunogenetics 24:146. (IP)
Baldwin TA and Hogquist KA. 2007. J. Immunol. 179:837.
Pascal V, et al. 2007. J. Immunol. 179:1751.
Burman AC, et al. 2007. Blood 110:1064.
Kincaid EZ, et al. 2007. J. Immunol. 179:3187.
Phan TG, et al. 2007. Nature Immunol. 8:992.
Nakano-Yokomizo T, et al. 2011. J. Exp Med. 208:1661. PubMed
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