Download 2281-MC-025 Bax 6A7 for pdf

TREVIGEN Product Data
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For Research Use Only. Not For Use In Diagnostic Procedures.
Procedure for Immunoblotting using Peroxidase Detection:
Anti-Bax YTH-6A7 Monoclonal Antibody
Antibody blocking solution: 5% (v/v) bovine serum in PBS.
Size: 25 µg
Catalog #: 2281-MC-025
Description: The Bcl-2 family of proteins plays a crucial role in the regulation of cell death in
many eukaryotic systems. Bax has been shown to redistribute from the cytosol to the
mitochondria during apoptosis, and overexpression of Bax can accelerate cell death.
Coregulation of Bax dimer formation and intracellular localization are associated with Bax
conformational changes. Anti-Bax 6A7 is conformation dependent and will recognize detergent
treated but not native Bax. When used in conjunction with the anti-Bax YTH-5B7 (Cat# 2280MC-100), which recognizes native Bax, the researcher may selectively identify the activated
form of Bax.
Transfer the electrophoresed proteins to a PVDF membrane following manufacturer’s
recommendations. Incubate the membrane for 1/2 hour at room temperature in antibody
blocking solution.
Incubate the membrane for 1 - 4 hours at room temperature in antibody blocking solution
containing a 1:1000 dilution of anti-Bax YTH-6A7 antibody. Empirical determination of
primary antibody concentration will be required for optimal results.
Wash the membrane at room temperature for 15 minutes with 3 changes of 0.05% Tween
in PBS. Changing the membrane containers often reduces background.
Physical State: This antibody is provided as purified immunoglobulin from mouse ascites in
1X PBS containing 0.01% sodium azide.
Incubate the membrane at room temperature for 1 hour in antibody blocking solution containing diluted anti-mouse IgG HRP. Empirical determination of secondary antibody concentration will be required for optimal results.
Immunogen: A synthetic peptide corresponding to amino acids 12 to 24 of the human Bax
sequence.
Wash the membrane for 15 minutes with 3 changes of 0.05% Tween in PBS.
Ig Class: IgG1
Develop peroxidase reaction using e.g. chemiluminescence (Trevigen’s PeroxyGlow A,
Cat# 4855-20-13, and PeroxyGlow B, Cat# 4855-20-14).
Specificity: Anti-Bax YTH-6A7 cross reacts with human, mouse, and rat Bax.
Storage: Store at 4˚C.
Applications: Western analysis, immunoprecipitation, and immunopurification. For western
blots, an antibody dilution of 1:1000 is recommended.
References: 1.
Suzuki, M., R.J. Youle, and N. Tjandra. 2000. Structure of bax. Coregulation of dimer formation and
intracellular localization.Cell 103:645-54.
2. Hsu, Y.-T., K.G. Wolter, and R.J. Youle. 1997. Cytosol-to-membrane redistribution of Bax and Bcl-XL
during apoptosis. Proc. Natl. Acad. Sci. USA 94:3668-3672.
3. Hsu, Y.-T., and R.J. Youle. 1997 Nonionic detergents induce dimerization among members of the
Bcl-2 family. J. Biol. Chem. 272:13829-13834.
4. Neuchushtan, A., C.L. Smith, Y.-T. Hsu, and R.J. Youle. 1999. Conformation of the Bax C-terminus
regulates subcellular location and cell death. EMBO J. 18:2330-2341
21 kDa
Western Blot analysis of murine WEHI cells. Cell
lysate from approximately 106 cells was loaded onto
a 12% SDS-PAGE gel. Proteins were separated by
electrophoresis and transferred to PVDF membrane. The membrane was incubated in antibody
blocking solution containing 1 µg/ml anti-Bax YTH6A7 monoclonal antibody, followed by chemiluminescent detection using an anti-mouse secondary
antibody conjugate .
© 2001 Trevigen, Inc. All Rights Reserved. Trevigen is a registered trademark of Trevigen, Inc.
TREVIGEN
Anti-Bax YTH-6A7
Monoclonal Antibody
v10705
Catalog #: 2281-MC-025
Storage: 4˚C
®
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