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 nd EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml. Label: FITC State: Liquid purified Ig fraction Label: Fluorescein isothiocyanate isomer 1
Concentration	lot specific
Conjugation	FITC
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 – 0.5 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. (It is recommended that the tubes are protected from light, since most fluorochromes are light sensitive.) 7. Wash 2 times at 4°C. 8. Resuspend the cell pellet in 50 μl ice cold media B. 9. 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: FITC Rat IgG1 Percentage of cells stained above control: Mesenteric Lymph Node Cells 95%
Reference Data

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