CD44 antibody

Catalog No. ABIN114366

Product Details anti-CD44 Antibody

Target: CD44

Reactivity: Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD44 antibody is un-conjugated

Application: Flow Cytometry (FACS), Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Immunohistochemistry (Frozen Sections) (IHC (fro))

Specificity: Reacts with rat CD44 (Pgp-1). This antibody recognizes an epitope on both standard CD44 and its splice variant.

Purification: Protein G affinity chromatography.

Immunogen: T cell blasts.

Clone: OX-49

Isotype: IgG2a

Application Details

Application Notes: This antibody is suitable for Immunoprecipitation, Flow cytometry (See protocol), WesternBlotting and Immunohistochemisty on frozen and paraffin sections.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with 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 thissuspension to each tube (each tube will then contain 1x10e6 cells, representing 1 test). 4. To each tube, add 1. 0-0. 5 µg* of ABIN114366 or ABIN114366X. 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 (FITC Goat anti-mouse IgG (H+L)) at 1: 500 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 arelight 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 propidiumiodide 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 (100µl of 2M sodium azide in 100 mls). Results-Tissue Distribution: Rat Strain: WistarCell Concentration: 1 x 10e6 cells per testsAntibody Concentration Used: 0. 2 µg/10e6 cellsIsotypic Control: Mouse IgG2a. Cell Source-Percentage of cells stained above control: Thymus: 82. 1%Spleen: 53. 5%Lymph Node: 87. 1%N. B. Appropriate control samples should always be included in any labelling studies. * For optimal results in various applications, it is recommended that each investigatordetermine dilutions appropriate for individual use.

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS buffer with 0.02 % sodium azide as preservative.

Preservative: Sodium azide

Precaution of Use: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.

Target Details for CD44

Target: CD44

Alternative Name: CD44 (CD44 Products)

Background: This antigen is expressed on most leukocytes (except a sub population of B cells) and increases upon activation. This antibody binds extracellularly to the standard (S) form on rat leukocytes, but it is not known if they bind to the N-terminal region. It has also been reported that the antibody may bind to melanoma cell lines that express CD44V (splice variant form). CD44 is expressed on most leukocytes except a sub population of B cells. Its expression is increased on T and B blasts.Synonyms: CDw44, ECMR-III, Epican, Extracellular matrix receptor III, GP90 lymphocyte homing/adhesion receptor, HUTCH-I, Heparan sulfate proteoglycan, Hermes antigen, Hyaluronate receptor, LHR, MDU2, MDU3, MIC4, PGP-1, Phagocytic glycoprotein 1

Gene ID: 10116

UniProt: P26051

Pathways: Glycosaminoglycan Metabolic Process, Autophagy, Negative Regulation of intrinsic apoptotic Signaling