TRAF1 antibody (AA 1-410)

Catalog No. ABIN5541248

This Mouse Monoclonal antibody specifically detects TRAF1 in WB. It exhibits reactivity toward Mouse.

Quick Overview for TRAF1 antibody (AA 1-410) (ABIN5541248)

Target: TRAF1 (TNF Receptor-Associated Factor 1 (TRAF1))

Reactivity: Mouse

Host: Mouse

Clonality: Monoclonal

Conjugate: This TRAF1 antibody is un-conjugated

Application: Western Blotting (WB)

Clone: 3D4

Product Details anti-TRAF1 Antibody

Binding Specificity: AA 1-410

Specificity: This antibody reacts with mouse TRAF1 (56 kDa) on Western blotting. This antibody doesn't react with mouse TRAF2, TRAF3, TRAF4, TRAF5, and TRAF6.

Cross-Reactivity (Details): Does not react with Human (Jurkat, U937).

Purification: Protein A agarose

Immunogen: Recombinant full-length mouse TRAF1 (1-410 aa)

Isotype: IgG1

Application Details

Application Notes: Western blot: 1 μg/mL for chemiluminescence detection system.

Protocol: SDS-PAGE & Western Blotting 1) Wash the cells 3 times with PBS and suspend with 10 volume 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 cold Lysis buffer to make 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 manufact ure's manual for precise 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 suggest in the APPLICATIONS for 1 hour at room temperature. (The concentration of antibody will depend on condition.) 8) Wash the membrane with PBS-T [0.05 % Tween-20 in PBS] (5 minutes x 3 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 (10 minutes x 3 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriate chemiluminescence reagent for 1 minute. 12) Remove extra reagent from the membrane by dabbing with paper towel, and seal it in plastic wrap. 13) Expose to an X-ray film in a dark room for 3 minutes. 14) Develop the film as usual. The condition for exposure and development may vary. (Positive controls for Western blotting mouse kidney, mouse liver)

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: PBS containing 50 % glycerol, pH 7.2. No preservative is contained.

Preservative: Azide free

Storage: -20 °C

Storage Comment: Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.

Target Details for TRAF1

Target: TRAF1 (TNF Receptor-Associated Factor 1 (TRAF1))

Alternative Name: traf1,ebi6

Background: Tumor necrosis factor receptor (TNF-R) superfamily such as TNFR1, TNFR2, CD30 and CD40-mediated signaling is provided by the TRAF (TNF receptor associated factors) family. The members of the TRAF family form either homodimers or heterodimers result in extensive diversity and specificity in their signal transduction pathways. TRAF1 and TRAF2 were found to associate with the cytoplas mic domains of the TNFR2. However, TRAF1 is indirec tly associated TNFR2 via dimerization with TRAF2. TRAF1 and TRAF2 bind independently to the same region of CD30. Both TRAF1 and TRAF2 interact strongly with A20, a TRAF2-mediated NF- κ B activation indirectly through the formation of a TRAF2-TRAF1-A20 complex.

UniProt: P39428

Pathways: NF-kappaB Signaling, Apoptosis, Cell-Cell Junction Organization, Asymmetric Protein Localization