BID anticorps (AA 61-118)

N° du produit ABIN5540048

Cet anticorps Souris Monoclonal détecte spécifiquement BID dans WB. Il présente une réactivité envers Humain.

Aperçu rapide pour BID anticorps (AA 61-118) (ABIN5540048)

Antigène: BID (BH3 Interacting Domain Death Agonist (BID))

Reactivité: Humain

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp BID est non-conjugé

Application: Western Blotting (WB)

Clone: 5C9

Détail du produit anti-BID anticorps

Épitope: AA 61-118

Specificité: This antibody reacts with BID.

Réactivité croisée (Details): Does not work with Mouse (WR19L, L5178Y, NIH/3T3) and Rat (Rat-1 BHK).

Purification: Protein A agarose

Immunogène: Recombinant human BID (61-118 aa)

Isotype: IgG1

Information d'application

Indications d'application: Western blot: 1-5 μg/mL for chemiluminescence detection system. For details see protocol below.

Protocole: 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 2 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 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 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 on the membrane, then incubate it with appropriate chemilumi nescence reagent for 1 minute. Remove extra reagent from the membrane by dabbing with paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. The condition for exposure and development may vary. (Positive controls for Western blotting ZR-75-1, U937, HL-60)

Restrictions: For Research Use only

Stockage

Format: Liquid

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

Agent conservateur: Azide free

Stock: -20 °C

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

Détails sur BID

Antigène: BID (BH3 Interacting Domain Death Agonist (BID))

Autre désignation: bid

Sujet: Apoptosis is a major form of cell death characterized by several morphological features that include chromatin condensation and fragmentation, cell membrane blebbing, and formation of apoptotic bodies. BID is an apoptosis promoting protein, which possesses only BH3 domain that is a common structure of Bcl-2 family members. Once full length BID (p22) is cleaved with caspases, cleaved BID (p 15) induces a conformational change and oligomerization of a protein named BAK on mitochondrial membrane. It is thought that oligomerized BAK contribute to forming pore s that release cytochrome c into cytosol.

UniProt: P55957

Pathways: Apoptose, Caspase Cascade in Apoptosis, Positive Regulation of Endopeptidase Activity