Activated Leukocyte Cell Adhesion Molecule (ALCAM) Antikörper (FITC)

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

Konjugat: FITC

Applikation: Durchflusszytometrie (FACS)

Produktnummer: ABIN131590

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 myelo ma cell P3x63/Ag8 with Balb/c mouse splenocyte immunized with human thymic epithelial cells.

Format: Liquid

Isotyp: IgG1

Klon: 3A6

Spezifität: This antibody reacts with human CD166 antigen.

Anwendungen

Applikationshinweise: Flow cytometric analysis for floating cells . Protocol 1: We 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 washing buffer (5x10 6 cells/mL). 3) Add 50 µL of the cell suspension into each tube, and centrifuge at 500xg for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 4) Add 10 µL of normal goat serum containing 1 mg/mL normal human IgG and 0.1% NaN 3 to the cell pellet after tapping. Mix well, and incubate for 5 minutes at room temperature (20~25 o C). 5) Add 30 µL of the FITC labeled CD166 monoclonal antibody (10-20 µg/mL) diluted with the washing buffer. Mix well, and incubate for 30 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) Resuspend the cells with 500 µL 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, 1 mg/mL normal human IgG and 0.1% NaN 3 (5x10 6 cells/mL). 3) Add 20 µL of the non diluted FITC labeled CD166 monoclonal antibody (50 µg/mL) into each tube. 4) Add 50 µL 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 for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 6) Resuspend the cells with 500 µL of the washing buffer and analyze by a flow cytometer. Flow cytometric analysis for whole blood cells We usually use Falcon tubes or equivalents as reaction tubes for all step described below. 1) Add 20 µL of the non diluted FITC labeled CD166 monoclonal antibody (50 µg/mL) into each tube. 2) Add 50 µL of whole blood into each tube. Mix well, and incubate for 30 minutes at room temperature (20~25 o C). 3) Lyse with OptiLyse C (for analysis on Beckman Coulter instruments) or OptiLyse B (for analysis on BD instruments), using the procedure recommended in the respective package inserts. 4) Resuspend the cells with 500 µL of the washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN 3 ] and analyze by a flow cytometer. Positive control for flow cytometry: Jurkat cells Flow cytometry: 10-20 µg/mL (final concentration) Research Applications.

Buffer: 50 µg IgG in 1 mL volume of PBS containing 1% BSA and 0.1% NaN 3 . * Azide may react with copper or lead in plumbing system to form explosive metal azides. Therefore, always flush plenty of water when disposing materials containing azide into drain.

Forschungsgebiet: CD Antigene

Beschränkungen: Nur für Forschungszwecke einsetzbar

Publikationen

Publikationen: Lehner, OFarrell: "Expression and function of Drosophila cyclin A during embryonic cell cycle progression." in: Cell, Vol. 56, Issue 6, pp. 957-68, 1989 (PubMed).

Tsai-Pflugfelder, Liu, Liu et al.: "Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 85, Issue 19, pp. 7177-81, 1988 (PubMed).

Parham, Brodsky: "Partial purification and some properties of BB7.2. A cytotoxic monoclonal antibody with specificity for HLA-A2 and a variant of HLA-A28." in: Human immunology, Vol. 3, Issue 4, pp. 277-99, 1982 (PubMed).

Burbelo, Hall: "14-3-3 proteins. Hot numbers in signal transduction." in: Current biology : CB, Vol. 5, Issue 2, pp. 95-6, 1995 (PubMed).

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).