bax抗体使用说明

/products/

Bax Antibody #2772

PhosphoSitePlus

?

protein, site, and accession data:

Bax

No.

Size

Price

2772S

100 ul ( 10 western blots )

contact

cst_intsales@

2772

carrier free & custom formulation / quantity

email request

Matching Secondary Antibody

Applications

Reactivity

Sensitivity

MW (kDa)

Source

W IP

H M R Mk

Endogenous

20

Rabbit

Applications Key:

W=Western Blotting

IP=Immunoprecipitation

Reactivity Key:

H=Human

M=Mouse

R=Rat

Mk=Monkey

Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on

100% sequence homology.

Protocols

2772:

Immunoprecipitation

,

Western Blotting

Specificity / Sensitivity

Bax Antibody detects endogenous levels of total Bax protein. The antibody does not cross- react

with other Bcl-2 family members.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding

to the amino-terminal residues of human Bax. Antibodies are purified by protein A and peptide

affinity chromatography.

Western Blotting

Western blot analysis of extracts from 293 (human), COS (monkey), L929 (mouse) and PC12

(rat) cells, using Bax Antibody.

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence

?

Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence

?

Bax siRNA I (+) or

SignalSilence

?

Bax siRNA II (+), using Bax Antibody and α

-Tubulin (11H10) Rabbit mAb

#2125. The Bax Antibody confirms silencing of Bax expression and α

-Tubulin (11H10) Rabbit

mAb is used to control for loading and specificity of Bax siRNA.

IP

Immunoprecipitation of Bax from Jurkat cell extract, using Bax Antibody. Lane 1 is the lysate

control; lane 2 is antibody alone; lane 3 is antibody plus lysate.

Background

Bax is a key component for cellular induced apoptosis through mitochondrial stress (1). Upon

apoptotic stimulation, Bax forms oligomers and translocates from the cytosol to the mitochondrial

membrane (2). Through interactions with pore proteins on the mitochondrial membrane, Bax

increases the membrane's permeability, which leads to the release of cytochrome c from

mitochondria, activation of caspase-9 and initiation of the caspase activation pathway for

apoptosis (3,4).

1.

Wei, M.C. et al. (2001)

Science

292, 727-730.

2.

Jurgensmeier, J.M. et al. (1998)

Proc. Natl. Acad. Sci. USA

95, 4997-5002.

3.

Narita, M. et al. (1998)

Proc. Natl. Acad. Sci. USA

95, 14681-14686.

4.

Marzo, I. et al. (1998)

Science

281, 2027-2031.

Application References

?

Xie, S. et al. (2001) Reactive oxygen species-induced phosphorylation of p53 on serine 20

is mediated in part by Polo-like kinase-3.

J. Biol. Chem.

276 (39), 36194-36199.

Applications:

Western Blotting

?

Yusta, B. et al. (2002)

J Biol Chem

277, 24896-906.

Applications:

Western Blotting

?

Leu, J. I. et al. (2004) Mitochondrial p53 activates Bak and causes disruption of a Bak-Mcl1

complex.

Nat. Cell. Biol.

6, 443-450.

Applications:

IP Western Blotting

?

Cheong, J. W. et al. (2003) Induction of apoptosis by apicidin, a histone deacetylase

inhibitor, via the activation of mitochondria-dependent caspase cascades in human

Bcr-Abl-positive leukemia cells.

Clin. Cancer Res.

9, 5018-5027.

Applications:

Western Blotting

?

Rusinol, A. E. et al. (2004) AKT/protein kinase B regulation of BCL family members during

oxysterol- induced apoptosis.

J. Biol. Chem.

279, 1392-1399.

Applications:

Western

Blotting

?

Widenmaier, S.B. et al. (2009)

J Biol Chem

284, 30372-82.

Applications:

Western

Blotting

?

Ikner, A. and Ashkenazi, A. (2011)

J Biol Chem

286, 21546-54.

Applications:

Western

Blotting

?

Pritchard, J.R. et al. (2011)

Cancer Res

71, 5850-8.

Applications:

Western Blotting

Have

you

published research involving the use of our products? If so we'd love to hear about it.

Please

let us know

!

Companion Products

?

6321 SignalSilence

?

Bax siRNA I

?

2762 Bcl-xL Antibody

?

4592 Bik Antibody

?

9292 Bad Antibody

?

7074 Anti-rabbit IgG, HRP-linked Antibody

?

7720 Prestained Protein Marker, Broad Range (Premixed Format)

?

7727 Biotinylated Protein Ladder Detection Pack

?

7003 20X LumiGLO

?

Reagent and 20X Peroxide

For Research Use Only. Not For Use In Diagnostic Procedures.

For Western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS,

0.1% Tween-20 at 4°

C with gentle shaking, overnight.

Products available from Cell Signaling Technology are linked by their respective catalog numbers.

A. Solutions and Reagents

NOTE:

Prepare solutions with Milli-Q or equivalently purified water.

1.

1X Phosphate Buffered Saline (PBS).

2.

1X SDS Sample Buffer:

(

#7722

,

#7723

) 62.5 mM Tris-HCl (pH 6.8 at 25°

C), 2% w/v

SDS, 10% glycerol, 50 mM DTT, 0.01% w/v bromophenol blue or phenol red.

3.

Transfer Buffer:

25 mM Tris base, 0.2 M glycine, 20% methanol (pH 8.5).

4.

10X Tris Buffered Saline (TBS):

(

#9997

) To prepare 1 liter of 10X TBS: 24.2 g Tris

base, 80 g NaCl; adjust pH to 7.6 with HCl (use at 1X).

5.

Nonfat Dry Milk:

(

#9999

) (weight to volume [w/v]).

6.

Blocking Buffer:

1X TBS, 0.1% Tween-20 with 5% w/v nonfat dry milk; for 150 ml,

add 15 ml 10X TBS to 135 ml water, mix. Add 7.5 g nonfat dry milk and mix well. While

stirring, add 0.15 ml Tween-20 (100%).

7.

Wash Buffer:

1X TBS, 0.1% Tween-20 (TBS/T).

8.

Bovine Serum Albumin (BSA):

(

#9998

).

9.

Primary Antibody Dilution Buffer:

1X TBS, 0.1% Tween-20 with 5% BSA; for 20 ml,

add 2 ml 10X TBS to 18 ml water, mix. Add 1.0 g BSA and mix well. While stirring, add

20 μl Tween

-20 (100%).

10.

Phototope

?

-HRP Western Blot Detection System:

(

#7071 anti-rabbit

) or (

#7072

anti- mouse

) Includes biotinylated protein ladder, secondary (

#7074 anti- rabbit

) or