Il2ra Mouse Monoclonal Antibody [Clone ID: OX-39]

Specifications

Product Data
Clone Name	OX-39
Applications	ELISA, FC, IHC
Recommend Dilution	Flow cytometry. Immunohistochemistry on frozen and paraffin sections. ELISA.
Reactivity	Rat
Host	Mouse
Clonality	Monoclonal
Immunogen	T blasts from a mixed lymphocyte reaction between purified CD4 positive T cells and irradiated spleens. Donor: BALB/c spleen Fusion Partner: NSO/1
Specificity	This monoclonal antibody recognizes the smaller (alpha subunit) 55kD chain of the IL-2 receptor found on activated rat T cells, thymic dendritic cells but not resting lymphocytes. This antibody binds to the rat interleukin-2 receptor designated CD25 and has proven to be an important marker for activated T cells (2).
Formulation	PBS and 0.02% NaN3 State: Purified State: Liquid purified Ig
Concentration	lot specific
Purification	Protein G Chromatography
Conjugation	Unconjugated
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 subunit alpha
Database Link	Entrez Gene 25704 Rat UniProt ID P26897
Background	The Interleukin 2 Receptor alpha and beta chains, together with the common gamma chain, constitute the high affinity IL2 receptor present on activated T and B cells, thymocyte subset, pre B cells and T regulatory cells. Homodimeric alpha chains result in low affinity receptor, while homodimeric beta chains produce a medium affinity receptor. Normally an integral membrane protein, soluble IL2 Receptor alpha has been isolated and determined to result from extracellular proteolysis. Alternately spliced IL2 Receptor alpha mRNAs have been isolated, but the significance of each is presently unknown.
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®- Rat 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 1x10e6 cells, representing 1 test). 4. To each tube, add 0.25 - 0.5 µg* of this Ab. 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 (PE Goat anti-mouse IgG (H+L)) at 1:20 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes are protected from light since most fluorochromes are light sensitive). 10. Wash 2 times at 4°C in media B. 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: Rat Strain: Wistar Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.25 µg/10e6 cells Isotypic Control: Mouse IgG1, kappa Cell Source Percentage of cells stained above control: Thymus - unactivated: 0% Con-A activated thymocytes: 88.9%
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.