Cd44 Rat Monoclonal Antibody [Clone ID: KM81]

Specifications

Product Data
Clone Name	KM81
Applications	FC, IHC
Recommend Dilution	Flow Cytometry Analysis (see Protocol).
Reactivity	Mouse
Host	Rat
Clonality	Monoclonal
Immunogen	Bone Marrow Derived Stromal Cells (clone BMS2). Donor: Lou/MN Rat. Fusion Partner: SP 2/0.
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 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).
Formulation	PBS containing 0.02% Sodium Azide as preservative. State: Purified State: Liquid purified Ig fraction.
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	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 1x10e6 cells, representing 1 test). 4. To each tube, add 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 (FITC Goat anti-rat 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 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: Mouse Strain: BALB/c Cell Concentration: 1x10e6 cells per test Antibody Concentration Used: 0.5 µg/10e6 cells Isotypic Control: 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
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.