FADD Antikörper (AA 1-208)

Produktnummer ABIN5541517

Dieses Anti-FADD-Antikörper ist ein Maus Monoklonal-Antikörper zur Detektion von FADD in WB. Geeignet für Human.

Kurzübersicht für FADD Antikörper (AA 1-208) (ABIN5541517)

Target: FADD (Fas (TNFRSF6)-Associated Via Death Domain (FADD))

Reaktivität: Human

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser FADD Antikörper ist unkonjugiert

Applikation: Western Blotting (WB)

Klon: 4G3

Produktdetails anti-FADD Antikörper

Bindungsspezifität: AA 1-208

Spezifität: This antibody reacts with human FADD. Occasionally doublet bands can be seen.

Kreuzreaktivität (Details): Does not react with Mouse (NIH/3T3, WR19L, BaF/3, P19), and Rat (PC12).

Aufreinigung: Protein A agarose

Immunogen: Recombinant full-length human FADD (1-208 a.a.)

Isotyp: IgG1

Anwendungsinformationen

Applikationshinweise: Western blot : 1 μg/mL for chemiluminescence detection system. For details see protocol below.

Protokoll: SDS-PAGE & Western blotting 1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1 % NP-40, 2 mM EDTA, 10 % glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the Lysis buffer to make an 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli's sample buffer. 4) Boil the samples for 3 minutes and centrifuge. Load 10 μ L of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi-dry transfer system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20 % MeOH). See the manufacture's manual for specific transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 o C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7.2 containing 1 % skimmed milk as suggested in the APPLICATIONS for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental conditions.) 8) Wash the membrane with PBS-T [0.05 % Tween-20 in PBS] (5 minutes x 6 times). 9) Incubate the membrane with the 1:10,000 HRP-conjugated anti-mouse IgG diluted with 1 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS-T (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriate chemilumi nescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 1 minute. Develop the film as usual. The conditions for exposure and development may vary. (Positive controls for Western blotting HeLa, MRC-5, ZR-75-1, A431, HEp-2)

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Lyophilized

Buffer: PBS ( pH 7.2) / 1 % Sucrose. Contains no preservatives.

Konservierungsmittel: Without preservative

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Prior to reconstitution store at 2-8°C. Following reconstitution store at -20°C. Avoid repeated freezing and thawing. Shelf life: one year from despatch.

Haltbarkeit: 12 months

Details zu FADD

Target: FADD (Fas (TNFRSF6)-Associated Via Death Domain (FADD))

Andere Bezeichnung: fadd

Hintergrund: FADD (Fas-associated death domain protein)/MORT1 has been identified as a protein that associates specifically with the intracellular domain of Fas. It contains both a death doma in (DD) and a death effecter domain (DED). DD is found in several death-inducing receptors of the TNF family, including Fas (CD95/APO-1) and TNFR-1. DED is found at the N-terminus of FADD and it is also present within the ICE-like protease, caspase-8 (FLICE/Mch5/MACH) and caspase-10 (FLICE2). Both DD and DED are able to associate with homologous regions in other proteins, and thus prompt binding of such proteins to one another. Upon activation, Fas relays death signals through DD, which interacts with the DD of the adaptor molecules FADD, recruiting them to the membrane. FADD then associates with caspase-8 through DED, leading to the assembly of a death-inducing signaling complex (DISC). DISC-associated caspase-8 subsequently initiates proteolytic activation of other caspase, which in turn leads to apoptosis.

UniProt: Q13158

Pathways: Apoptose, TLR Signalweg, Activation of Innate immune Response, Positive Regulation of Endopeptidase Activity, Toll-Like Receptors Cascades