Cd44 Rat Monoclonal Antibody [Clone ID: KM81]

CAT#: CL023R

Cd44 rat monoclonal antibody, clone KM81, PE

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Specifications

Product Data

• Clone Name: KM81
• Applications: FC
• Recommended Dilution: Flow cytometry.
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Immunogen: Bone Marrow Derived Stromal cells (clone BMS2).
  Donor: Lou/MN Rat.
  Fusion Partner: SP2/0.
• 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 on 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).
• Formulation: PBS, 0.02% NaN3 and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: PE
  State: Liquid
• Concentration: lot specific
• Purification: Protein G Chromatography
• Conjugation: PE
• Storage: Store the antibody undiluted at 2-8°C.
  DO NOT FREEZE!
  This product is photosensitive and should be protected from light.
• Stability: Shelf life: one year from despatch.
• Gene Name: CD44 antigen
• Database Link:
  Entrez Gene 12505 Mouse
  UniProt ID P15379
• 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: LHR, MDU2, MDU3, MIC4, CDw44, Epican, ECMR-III, HUTCH-I, Heparan sulfate proteoglycan, Hermes antigen, Hyaluronate receptor, PGP-1
• 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 1.0 - 0.5 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. (It is recommended that the tubes are protected from light, since most fluorochromes are light sensitive.)
  7. Wash 2 times at 4°C.
  8. Resuspend the cell pellet in 50 µl ice cold media B.
  9. 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:
  Mouse Strain: BALB/c
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 0.5 µg/10e6 cells
  Isotypic Control: PE Rat IgG2a
  
  Results - Strain Distribution:
  Cell Concentration: 1x10e6 cells per test
  Antibody Concentration Used: 0.5 µg /10e6 cells
  Strains Tested: BALB/c, CBA/J, C3H/He, C57BL/6, SWR
  Positive: BALB/c, CBA/J, C3H/He, C57BL/6, SWR
  Negative: none