|Validated Species:||Human, Mouse, Rat|
|Application:||Flow Cytometry, IHC-F, ICC, WB|
Data & Images
|Product Name||Anti-Bax Antibody|
|Description||Rabbit IgG polyclonal antibody for Apoptosis regulator BAX(BAX) detection. Tested with WB, IHC-F, ICC, FCM in Human;Mouse;Rat.|
|Cite This Product||Anti-Bax Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1013-1)|
|Replacement Item||This antibody may replace the following items: sc-6236|sc-526|sc-7480|sc-80658|sc-65532|sc-493|sc-70408|sc-70405|sc-23959|sc-70407|sc-20067|sc-526-G|sc-526-X from Santa Cruz Biotechnology.|
|Validated Species||Human, Mouse, Rat|
*This antibody is predicted to react with the above species based on antigen sequence similarities. Our Boster Guarantee covers the use of this product with the above species.
|Application||Flow Cytometry, IHC-F, ICC, WB
*Our Boster Guarantee covers the use of this product in the above tested applications.
**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.
|Recommended Detection Systems||Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(F) and ICC.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2018!
|Immunogen||A synthetic peptide corresponding to a sequence at the N-terminus of human Bax(19-33aa IMKTGALLLQGFIQD), different from the related mouse and rat sequences by one amino acid.|
|Cross Reactivity||No cross reactivity with other proteins|
|Contents||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Concentration||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Storage||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Purification||Immunogen affinity purified.|
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
|Protein Name||Apoptosis regulator BAX|
|Molecular Weight||21184 MW|
|Protein Function||Accelerates programmed cell death by binding to, and antagonizing the apoptosis repressor BCL2 or its adenovirus homolog E1B 19k protein. Under stress conditions, undergoes a conformation change that causes translocation to the mitochondrion membrane, leading to the release of cytochrome c that then triggers apoptosis. Promotes activation of CASP3, and thereby apoptosis. .|
|Tissue Specificity||Expressed in a wide variety of tissues. Isoform Psi is found in glial tumors. Isoform Alpha is expressed in spleen, breast, ovary, testis, colon and brain, and at low levels in skin and lung. Isoform Sigma is expressed in spleen, breast, ovary, testis, lung, colon, brain and at low levels in skin. Isoform Alpha and isoform Sigma are expressed in pro- myelocytic leukemia, histiocytic lymphoma, Burkitt's lymphoma, T- cell lymphoma, lymphoblastic leukemia, breast adenocarcinoma, ovary adenocarcinoma, prostate carcinoma, prostate adenocarcinoma, lung carcinoma, epidermoid carcinoma, small cell lung carcinoma and colon adenocarcinoma cell lines. .|
|Sequence Similarities||Belongs to the Bcl-2 family.|
|Subcellular Localization||Isoform Alpha: Mitochondrion membrane; Single-pass membrane protein. Cytoplasm. Colocalizes with 14- 3-3 proteins in the cytoplasm. Under stress conditions, undergoes a conformation change that causes release from JNK-phosphorylated 14-3-3 proteins and translocation to the mitochondrion membrane.|
|Alternative Names||Apoptosis regulator BAX;Bcl-2-like protein 4;Bcl2-L-4;BAX;BCL2L4;|
|Research Areas|||cell biology|apoptosis|intracellular|associated proteins| cell biology|bcl2 family| cancer|invasion/microenvironment|bcl 2 family| kits/ lysates/ other|elisa kits|apoptosis marker and proteins elisa kits| metabolism|pathways and processes|metabolism processes|cell death|apoptotic markers||
Background for Apoptosis regulator BAX
Dilution Ratios/Recommended Concentrations
At Boster we strive to provide the best Anti-Bax Antibody by testing all applications on non-spiked tissues and cell lines to ensure that the affinity of the antibody is enough to react to the endogenouse level of the target protein. Read more about our QC panel here.
|Recommended dilution ratios are listed below:|
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse|
Immunohistochemistry(Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
**Boster provides high sensitivity secondary antibody kits for Western blotting and IHC. For more info see Related Products below.
Anti-Bax Antibody Images
Click the images to enlarge.
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: Rat Testis Tissue Lysate
Lane 2: Rat Kidney Tissue Lysate
Lane 3: Rat Brain Tissue Lysate
Lane 4: Rat Ovary Tissue Lysate
Lane 5: HELA Cell Lysate
Lane 6: MM231 Cell Lysate
Lane 7: A549 Cell Lysate
Lane 8: JURKAT Cell Lysate
Lane 9: Human Placenta Tissue Lysate
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- Bax antigen affinity purified polyclonal antibody (Catalog # PA1013-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Bax at approximately 21&28KD. The expected band size for Bax is at 21KD.
Overlay histogram showing A431 cells stained with PA1013-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-BAX Antibody (PA1013-1,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,