Il2ra Rat Monoclonal Antibody [Clone ID: PC61.5.3]

Specifications

Product Data
Clone Name	PC61.5.3
Applications	FC, IHC, IP
Recommend Dilution	Immunoprecipitation. Immunohistochemistry on frozen sections. Flow Cytometry.
Reactivity	Mouse
Host	Rat
Clonality	Monoclonal
Immunogen	B6.1 CTL Cell line Donor: OFA rat spleen Fusion Partner: Mouse myeloma line P3X63Ag8.653
Specificity	Anti-mouse CD25 (IL-2R) monoclonal antibody reacts with the low affinity alpha chain of the interleukin-2 receptor antigen present on activated T and B cells in mice. CL8925B inhibits IL-2 binding and IL-2 dependent proliferation.
Formulation	PBS containing 0.02% sodium azide 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
Purification	Affinity chromatography on Protein G
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	interleukin 2 receptor, alpha chain
Database Link	Entrez Gene 16184 Mouse UniProt ID P01590
Synonyms	Interleukin-2 receptor alpha chain, IL-2 receptor alpha subunit, IL-2-RA, IL2-RA, p55, TAC antigen
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 0.2-0.1 μg* of AM26037BT-N or AM26037BT-L 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 tests Antibody Concentration Used: 0.1 μg/10e6 cells Isotypic Control: Biotin Rat IgG1 Cell Source Percentage of cells stained above control: Cell Blasts (Con A activated) 99.4% Thymus (unactivated) 1.7%
Reference Data

*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.