CD44 antibody (Biotin)

Catalog No. ABIN114363

Product Details anti-CD44 Antibody (Biotin)

Target: CD44

Reactivity: Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This CD44 antibody is conjugated to Biotin

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

Specificity: This monoclonal antibody recognizes rat CD44 (Pgp-1), also called CD44H. This antigen is expressed on most leukocytes (except a sub population of B cells) and increases upon activation. The OX-49 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).

Purification: Protein G Chromatography

Immunogen: T cell blasts. Donor: BALB/c spleen. Fusion Partner: myeloma cell line NSO/1.

Clone: OX-49

Isotype: IgG2a

Application Details

Application Notes: Flow Cytometry. Western Blotting. Immunohistochemistry 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 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 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 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 (Streptavidin-FITC) at 1: 500 dilution. 9. Incubate the tubes at 4°C for 30-60 minutes. (It is recommended that the tubes areprotected 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 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 testAntibody Concentration Used: 0. 5 µg/10e6 cellsIsotypic Control: Biotin Mouse IgG2aCell Source Percentage of cells stained above control:

Restrictions: For Research Use only

Handling

Concentration: 0.1 mg/mL

Buffer: PBS, 0.02 % NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/mL

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: CD44 is a type 1 transmembrane glycoprotein also known as Phagocytic Glycoprotein 1 (pgp 1) and HCAM. CD44 is the receptor for hyaluronate and exists as a large number of different isoforms due to alternative RNA splicing. The major isoform expressed on lymphocytes, myeloid cells, and erythrocytes is a glycosylated type 1 transmembrane protein. Other isoforms contain glycosaminoglycans and are expressed on hematopoietic and non hematopoietic cells. CD44 is involved in adhesion of leukocytes to endothelial cells, stromal cells, and the extracellular matrix.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