Activated Leukocyte Cell Adhesion Molecule (ALCAM) Antikörper

Antigen: Activated Leukocyte Cell Adhesion Molecule (ALCAM)

Synonyme: MEMD, CD166, FLJ38514, MGC71733, BEN, SC1, MuSC, AI853494, DM-GRASP, MGC27910, cd166, ALCAM, JC7, MGC83026, memd, MGC147089, DKFZp459H0124

Klonalität: Monoklonal (3A6)

Wirt: Maus

Reaktivität: Human

Applikation: Durchflusszytometrie (FACS)

Produktnummer: ABIN131589

Produktbeschreibung

Weitere Bezeichnung: CD166

Immunogen: This antibody was purified from hybridoma (clone 3A6) supernatant using protein-A Sepharose. This hybridoma was established by fusion of mouse myeloma cell P3x63/Ag8 with BALB/c mouse splenocyte immunized with human thymic epithelial cells.

Format: Liquid

Isotyp: IgG1  (Passende Sekundärantikörper)

Klon: 3A6

Spezifität: This antibody reacts with human CD166 antigen.

Anwendungen

Applikationshinweise: Flow cytometric analysis for floating cells . Protocol 1: We usually use Fisher tubes or equivalents as reaction tubes for all step described below. 1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN 3 ]. 2) Add 10 µ L of normal goat serum to the cell pellet after tapping. Mix well, and incubate for 5 minutes at room temperature (20~25 o C). 3) Add 30 µL of the CD166 monoclonal antibody (10-20 µ g/mL) diluted with the washing buffer. Mix well, and incubate for 30 minutes at room temperature (20~25 o C). 4) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 5) Add 30 µL of secondary antibody (1:40 FITC conjugated anti-mouse IgG/code no. IM-0819) diluted with the washing buffer. Mix well and incubate for 15 minutes at room temperature (20~25 o C). 6) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 7) Resespend the cells with 500 µ L of the washing buffer and analyze by a flow cytometer. . Protocol 2: We usually use Fisher tubes or equivalents as reaction tubes for all step described below. 1) Wash the cells 3 times with washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN 3 ]. 2) Resuspend the cells with PBS containing 25% normal goat serum and 0.1% NaN 3 (5x10 6 cells/ml). 3) Add 20 µ L of the CD166 monoclonal antibody (50 µ g/mL) diluted with the washing buffer into each tube. 4) Add 50 µ L of the cell suspension into each tube. Mix well, and incubate for 30 minutes at room temperature (20~25 o C). 5) Add 1 mL of the washing buffer followed by centrifugation at 500xg rpm for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 6) Resuspend the cells with 50 µ L of the washing buffer. 7) Add 20 µL of secondary antibody (1:10 FITC conjugated anti-mouse IgG/code no. IM-0819) diluted with the washing buffer into each tube. Mix well and incubate for 15 minutes at room temperature (20~25 o C). 8) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 9) Resuspend the cells with 500 µ L of the washing buffer and analyze by a flow cytometer Flow cytometry: 10-20 µ g/ml (final concentration) Research Applications.

Buffer: 100 µ g IgG in 100 µ L volume of PBS containing 50% glycerol. No preservative is contained.

Forschungsgebiet: CD Antigene

Beschränkungen: Nur für Forschungszwecke einsetzbar

Publikationen

Publikationen: Levesque, Heinly, Whichard et al.: "Cytokine-regulated expression of activated leukocyte cell adhesion molecule (CD166) on monocyte-lineage cells and in rheumatoid arthritis synovium." in: Arthritis and rheumatism, Vol. 41, Issue 12, pp. 2221-9, 1999 (PubMed).