Bax

Product Description for Bax

Mouse anti Mouse Bax 5B7.
Presentation: Azide Free
Product is tested for Enzyme Immunoassay, Immunocytochemistry/Immunofluorescence, Immunoprecipitation, Western blot / Immunoblot.

Properties for Bax

Product Category	Antibodies
Quantity	0.1 mg
Synonyms	Apoptosis regulator BAX, BCL2-associated X protein, BCL2L4, Bcl-2-like protein 4, Bcl2-L-4, FocusOn130, Review 004
Presentation	Azide Free
Reactivity	Ms
Applications	E, ICC/IF, IP, WB
Clonality	Monoclonal
Clone	5B7
Host	Mouse
Isotype	IgG1
Shipping to	Not USA/Canada
PDF datasheet	View Datasheet
Manufacturer	Acris Antibodies GmbH

Datasheet Extract

Immunogen	Swiss Prot Num: Q07813 Immunogen: Synthetic peptide (KLH conjugated) corresponding to amino acids 3–16 near the N-terminus of Mouse Bax. AA Sequence: `GSGEQLGSGGPTSS` Genename: 12028 Remarks: The immunogen sequence is not present in Human or Rat Bax. Hybridoma was established by fusion of mouse myeloma cell NS-1 with Balb/c mouse splenocyte.
Antibody type	Ab
Add. information	This product was originally produced by MBL International.
Application	ELISA. Western blotting: 1 µg/mL for chemiluminescence detection system. Immunofluorescence. Immunoprecipitation: 2 μg/200 μL of cell extract from 5x10e6 cells. Detailed procedure is provided in Protocols.
Background	Bax (Bcl-associated X protein) is a 21 kDa tumor suppressor protein that suppresses tumorigenesis and stimulates apoptosis in vivo. Bax has extensive amino acid homology to Bcl-2. It can homodimerize through its BH3 domain and it forms heterodimers with other Bcl-2 family members through its BH1 and BH2 domains. Overexpression of Bax promotes apoptosis and counters the death repressor activity of Bcl-2 and Bcl-xL. It is believed that the ratio of Bcl-2/Bax complexes to free protein controls the relative susceptibility of cells to death stimuli. Apoptotic stimuli cause the translocation of monomeric Bax from the cytosol to the mitochondria where it forms Bax homodimers. Localization of Bax to the mitochondria results in the activation of caspase-3, membrane blebbing, and nuclear fragmentation. Bax also induces mitochondrial dysfunction by increasing mitochondrial membrane permeability. It accelerates the opening of the mitochondrial porin channel VDAC, thus regulating the release of cytochrome c during apoptosis.
Concentration	1.0 mg/ml
Protocols	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°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°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 μL 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/cm2 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°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 POD-conjugated anti-mouse IgG diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at RT. 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. 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 3 minutes. Develop the film as usual. The condition for exposure and development may vary. Positive Controls: NIH/3T3, BaF/3, WR19L Immunoprecipitation 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°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°C and transfer the supernatant to another tube. 3) Add primary antibody as suggested in the APPLICATIONS into 200 µL of the supernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4°C. Add 20 µL of 50% protein A agarose resuspended in the cold Lysis buffer. Mix well and incubate with gentle agitation for 60 minutes at 4°C. 4) Wash the beads 3-5 times with the cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds). 5) Resuspend the beads in 20 μL of Laemmli’s sample buffer, boil for 3-5 minutes, and centrifuge for 5 minutes. Use 10 µL/lane for the SDS-PAGE analysis. (See SDS-PAGE & Western blotting.) Positive Contro: WR19L
General Readings	1) Hsu Y.T., et al. J. Biol. Chem. 272, 13829-13834 (1997). Clone 5B7 is used in this reference.
Storage	Store the antibody (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: one year from despatch.
Format	Purification: Protein-A Sepharose Chromatography of hybridoma supernatant. Buffer System: PBS, pH 7.2 containing 50% Glycerol without preservatives. State: Liquid purified IgG fraction. Azide Free
Species Reactivity	Species reactivity (tested): Mouse.
Specificity	Specificity: This antibody reacts with Mouse Bax (21 kDa) on Western blotting.
Gene ID	12028
Targets	Bax antibody

Accessory Products

Proteins and/or Positive Controls

Proteins for Bax (1 products)

Catalog No.	Species	Pres.	Purity	Source
PR27055-10	Bax
	Mouse	Purified	> 95 %	E. coli	1 µg / €240.00
					Neuromics Antibodies

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