CD45 / LCA Rat Monoclonal Antibody [Clone ID: IBL-5/25]

Specifications

Product Data
Clone Name	IBL-5/25
Applications	FC
Recommend Dilution	Immunohistochemistry on aceton-fixed frozen sections. Immunoprecipitation. Flow Cytometry.
Reactivity	Mouse
Host	Rat
Clonality	Monoclonal
Immunogen	IL-3 dependent mast cells derived from WB- +/+ mice Donor: Wistar spleen Fusion Partner: X63.653. Ag8
Specificity	Anti-mouse CD45 monoclonal antibody detects CD45 (L-CA) which is a transmembrane phosphotyrosine phosphatase expressed on leukocytes. This mAb induces the in vitro clustering of mouse lymphocytes (both T and B cells).
Formulation	PBS containing 0.02% sodium azide (NaN3) as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. Label: Biotin State: Liquid purified Ig fraction
Concentration	lot specific
Conjugation	Biotin
Storage Condition	Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing.
Gene Name	protein tyrosine phosphatase, receptor type, C
Database Link	Entrez Gene 19264 Mouse UniProt ID P06800
Synonyms	PTPRC, Leukocyte common antigen, L-CA, T200
Note	Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-M cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 μl of this suspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 2.0 μg of this antibody per 10e6 cells. 5. Vortex the tubes to ensure thorough mixing of antibody and cells. 6. Incubate the tubes for 30 minutes at 4°C. 7. Wash 2 times at 4°C. 8. Add 100 μl of secondary antibody (Streptavidin-FITC) at a 1/500 dilution. 9. Incubate tubes at 4°C for 30 - 60 minutes (It is recommended that tubes are protected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C. 11. Resuspend the cell pellet in 50 μl ice cold media B. 12. Transfer to suitable tubes for flow cytometric analysis containing 15 μl of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA. Media: A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + sodium azide (100 μl of 2M sodium azide in 100 mls). B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine serum albumin + sodium azide (100 μl of 2M sodium azide in 100 mls). Results: Tissue Distribution by Flow Cytometry Analysis: Mouse Strain: BALB/c Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 2.0μg/10e6 cells Isotypic Control: Biotin Rat IgG1 Percentage of cells stained above control: Mesenteric Lymph Node Cells 95%
Reference Data

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