CD44 (CD44) Antikörper

Antigen: CD44 (CD44)

Synonyme: IN, LHR, MC56, MDU2, MDU3, MIC4, Pgp1, CDW44, CSPG8, HCELL, MUTCH-I, ECMR-III, MGC10468

Klonalität: Monoklonal (5034-44-2)

Wirt: Maus

Reaktivität: Maus

Applikation: Zytotoxizitätstest (CyTox), Durchflusszytometrie (FACS)

Produktnummer: ABIN114216

Produktbeschreibung

Produktmerkmale: Synonyms: CDw44, PGP-I, ECMR-III, Hermes antigen, Epican, LHR, MDU2, MDU3, MIC4, Phagocyticglycoprotein I, Extracellular matrix receptor III, GP90 lymphocyte homing/adhesionreceptor, Hyaluronate receptor, Heparan sulfate proteoglycan

Weitere Bezeichnung: CD44 / PGP-1 / HUTCH-I

Swiss-Prot: P15379

Immunogen: B6 - Ly-1a spleenDONOR: BALB/c spleenFUSION PARTNER: P3-NS1-1-Ag4(NS1/1)

Format: Ascites

Isotyp: IgG2a  (Passende Sekundärantikörper)

Klon: 5034-44-2

Beschreibung: 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 ofdifferent isoforms due to alternative RNA splicing. The major isoform expressed onlymphocytes, myeloid cells, and erythrocytes is a glycosylated type 1 transmembraneprotein. Other isoforms contain glycosaminoglycans and are expressed on hematopoieticand non hematopoietic cells. CD44 is involved in adhesion of leukocytes to endothelialcells, stromal cells, and the extracellular matrix.

Spezifität: This monoclonal antibody recognizes the murine alloantigen Ly 24. 2. CD44 (Ly 24B. 2,Pgp-1. 2) is a 95 kDa glycoprotein previously known as phagocytic glycoprotein -1. It has awide tissue distribution and is found on bone marrow derived cells, lymphocytes and non -lymphoid tissue such as brain, liver, and kidney. There is variation in Ly 24 expressionbetween mice strains. Generally expression by Ly 24. 1 strains is high while in Ly 24. 2 it islower (2). The Ly 24 antigen is expressed by T lymphocytes during primary antigen stimulation. Thisantigen can be used as a marker to identify activated or memory T cells (3,4). Species: Mouse. Others not tested.

Anwendungen

Protokoll: FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare 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 cells to 1 x 10e6 cells in approximately 50 µl Media A in a microcentrifugetube (ie. 50 µl of cells resuspended to 2 x 10e7 cells / ml). (THE CONTENTS OF 1 TUBEREPRESENTS 1 TEST). 4. To each tube add 50 µl of 1/500 dilution of this Ab(final dilution 1/1000). 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 CLCC30201 (Goat anti-mouse IgG(H+L)-FITC conjugate)@ 1: 700. 9. Incubate 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 phosphate buffered saline. (This stains dead cells byintercalating 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 µlof 2M sodium azide in 100 mls). RESULTS - TISSUE DISTRIBUTION: PROCEDURE: as aboveANTIBODY CONCENTRATION: 1: 1000MOUSE STRAIN: C57BL/6CELL SOURCE: PERCENT STAININGThymus: 8. 8%Spleen: 36. 1%Lymph Node: 17. 9%

Applikationshinweise: Flow cytometry. (see Protocols)Method for Determining percent of positive cells in a poulation. (see Protocols)Method for Depleting a cells poulation of positive lymphocytes. (see Protocols)Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Konzentration: 1 x 10e6 cells per ml.Complement: Low-Tox®-M Rabbit ComplementComplement 1: 15Antibody 1: 1000CELL SOURCE: C.I.Spleen: 8Splenic T-cells: 36Lymph Node: 14Lymph Node T-cells: 21Thymus: 9PROCEDURE: Cells were treated as described in Recommended Method for

Lagerung: Prior to and following reconstitution store the antibody at -20°C. Avoid repeated freezing and thawing. Shelf life: one year from despatch.

Forschungsgebiet: CD Antigene, Oberflächenrezeptoren der Immunzellen

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

Mouse anti CD44 (HCAM) (Ly-24B.2) 5034-44.2

Publikationen

Publikationen: Sutton, Wijffels, Walker et al.: "Genetic and biochemical characterization of antigens encoded by the Ly-24 (Pgp-1) locus." in: Journal of immunogenetics, Vol. 14, Issue 1, pp. 43-57, 1987 (PubMed).

Butterfield, Fathman, Budd: "A subset of memory CD4+ helper T lymphocytes identified by expression of Pgp-1." in: The Journal of experimental medicine, Vol. 169, Issue 4, pp. 1461-6, 1989 (PubMed).

Lynch, Ceredig: "Mouse strain variation in Ly-24 (Pgp-1) expression by peripheral T cells and thymocytes: implications for T cell differentiation." in: European journal of immunology, Vol. 19, Issue 2, pp. 223-9, 1989 (PubMed).

Budd, Cerottini, Horvath et al.: "Distinction of virgin and memory T lymphocytes. Stable acquisition of the Pgp-1 glycoprotein concomitant with antigenic stimulation." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 138, Issue 10, pp. 3120-9, 1987 (PubMed).