CD6 (4F2) Antikörper

Antigen: CD6 (4F2)

Klonalität: Monoklonal (4F2)

Wirt: Maus

Reaktivität: Human

Applikation: Durchflusszytometrie (FACS)

Produktnummer: ABIN131584

Produktbeschreibung

Immunogen: This antibody was purified from hybridoma (clone 4F2) supernatant using protein-A Sepharose. This hybridoma was established by fusion of mouse myeloma cell P3x63/Ag8 with BALB/c mouse splenocyte immunized with COS7 cells transfected with CD6.

Format: Liquid

Isotyp: IgG1 kappa  (Passende Sekundärantikörper)

Klon: 4F2

Spezifität: This antibody reacts with human CD6 SRCR (Scavenger Receptor Cysteine Rich) domain 1.

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 CD6 (4F2 ) 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) Resuspend 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 CD6 (4F2) 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 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: Patel: "CD6." in: Journal of biological regulators and homeostatic agents, Vol. 14, Issue 3, pp. 234-6, 2001 (PubMed).