CD44 antibody (Biotin)

Catalog No. ABIN114206

The Rat Monoclonal anti-CD44 antibody has been validated for FACS and IHC (fro). It is suitable to detect CD44 in samples from Mouse.

Quick Overview for CD44 antibody (Biotin) (ABIN114206)

Target: CD44

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This CD44 antibody is conjugated to Biotin

Application: Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro))

Clone: KM81

Product Details anti-CD44 Antibody (Biotin)

Specificity: This monoclonal antibody recognizes a 95 kDa glycoprotein found on most hematopoietic cells (1). It is thought to be important in the regulation of migratory properties of lymphocytes during development and the regulation of the interaction with bone marrow stromal cells during hematopoiesis (2,3). CD44 functions as a receptor for hyaluronate, although some cells expressing CD44 do not bind hyaluronate (3,4). This antibody has been shown to inhibit the growth of lymphoid and myeloid cells in long term bone marrow cultures (3). It also blocks the adhesive interactions of B cell hybridomas to a cloned stromal line or to hyaluronate coated dishes (4).

Purification: Protein G Chromatography.

Immunogen: Bone Marrow Derived Stromal cells (clone BMS2). Donor: Lou/MN Rat. Fusion Partner: SP2/0.

Isotype: IgG2a

Application Details

Application Notes: Flow Cytometry. Immunohistochemistry.
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®-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 thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add 1. 0 µg* of this Ab 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 protectedfrom 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 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 DistributionMouse Strain: BALB/cCell Concentration: 1x10e6 cells per testAntibody Concentration Used: 1. 0 µg/10e6 cellsIsotypic Control: Biotin Rat IgG2aResults - Strain Distribution: Cell Concentration: 1x10e6 cells per testsAntibody Concentration Used: 1. 0 µg/10e6 cellsStrains Tested: BALB/c, CBA/J, C3H/He, C57BL/6, SWRPositive: BALB/c, CBA/J, C3H/He, C57BL/6, SWR

Restrictions: For Research Use only

Handling

Concentration: 0.1 mg/mL

Buffer: 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.

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

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: 12505

NCBI Accession: NP_001034240

UniProt: P15379

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