AM00283AF-N Bax antibody

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0.1 mg / €350.00
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Mouse anti Mouse Bax 5B7

AM00283AF-N

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  • AM00283AF-N

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
Target Category
Quantity 0.1 mg
Synonyms Apoptosis regulator BAX, BCL2-associated X protein, BCL2L4, Bcl-2-like protein 4, Bcl2-L-4
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
Material safety datasheet MSDS for Monoclonal Antibodies (de)

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
Isotype control AM33027AF-N
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

Accessory Products

Proteins and/or Positive Controls

Proteins for Bax (1 products)

Catalog No. Species Pres. Purity   Source  
PR27055-10

Bax

 BAX Structure and Sequence. Mouse Purified > 95 % E. coli
1 µg / €270.00
  Neuromics Antibodies
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