Product nameAnti-Bax antibody [E63]
See all Bax primary antibodies
DescriptionRabbit monoclonal [E63] to Bax
SpecificityExpression levels of BAX protein vary with sample type. Induction may be required if endogenous expression is low.
Tested applicationsSuitable for: IHC-P, WB, IP, Sandwich ELISAmore details
Unsuitable for: Flow Cyt or ICC
Species reactivityReacts with: Mouse, Rat, Human, Chinese hamster
Predicted to work with: Cow
- WB: Recombinant Human Bax protein (Tagged) (ab85157), HeLa cell lysate. IHC-P: Human lymph node and rat kidney tissues.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
- Anti-Cytochrome C antibody [37BA11] (ab110325)
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- ABT-737, BH3 mimetic Bcl-xL, Bcl-2 and Bcl-w inhibitor (ab141336)
- 2-Methoxyantimycin A3, Bcl-xL and Bcl-2 inhibitor (ab141523)
- Z36, Novel inhibitor of Bcl-XL (ab141757)
- Recombinant Human Bax protein (ab173026)
- Human BAX knockout HeLa cell lysate (ab263841)
sELISA pair antibody
Our Abpromise guarantee covers the use of ab32503 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||1/1000 - 1/10000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).|
|Sandwich ELISA||Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Mouse monoclonal [2D2] to Bax - BSA and Azide free (ab77566).|
FunctionAccelerates 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 specificityExpressed 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 similaritiesBelongs to the Bcl-2 family.
DomainIntact BH3 motif is required by BIK, BID, BAK, BAD and BAX for their pro-apoptotic activity and for their interaction with anti-apoptotic members of the Bcl-2 family.
Cellular localizationCytoplasm and Mitochondrion membrane. 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.
- Information by UniProt
- Apoptosis regulator BAX antibody
- BAX antibody
- Bax-protein antibody
Lane 1: Hap1 wildtype cell lysate (20 µg)
Lane 2: BAX Hap1 knockout cell lysate (20 µg)
Lane 3: HeLa wildtype cell lysate (20 µg)
Lane 4: BAX HeLa knockout cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32503 observed at 21 k Da kDa. Red - loading control, ab8245 observed at 37 kDa.
ab32503 was shown to react with Bax in HeLa wildtype. Loss of signal was observed when knockout sample ab263841 was used. Wild-type and Bax knockout samples were subjected to SDS-PAGE. ab32503 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Bax knockout HAP1 cell lysate (20 µg)
Lanes 1 - 2: Merged signal (red and green). Green - ab32503 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa or ab18058, observed at 130 kDa.
This western blot image is a comparison between ab32503 and a competitor's top cited rabbit polyclonal antibody.
Purified ab32503 staining Bax in Human lung carcinoma tissue section by immmunohistochemistry (IHC-P- Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraffin and heat mediated antigen retrieval was performed using EDTA buffer (pH 9.0). Samples were incubated with primary antibody at 1:500 dilution. A goat anti-rabbit IgG H&L (HRP) (ab97051) was used as a secondary antibody at 1:500 dilution. Cytoplasmic staining on human lung carcinoma.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, 10μg
Lane 2 (+): HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32503 in HeLa whole cell lysate
Purified ab32503 immunoprecipitating Bax in HeLa lysates. For western blotting, the primary antibody used was purified ab32503 at 1/1000 dilution. Ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection at 1/1000 dilution. Capture antibody was used at a 1/20 dilution. Blocking and diluting buffer used was 5% NFDM/TBST.
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Bax knockout HAP1 cell lysate (20 µg)
Lanes 1 - 2: Merged signal (red and green). Green - ab32503 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32503 was shown to recognize Bax when Bax knockout samples were used, along with additional cross-reactive bands. Wild-type and Bax knockout samples were subjected to SDS-PAGE. ab32503 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
IHC image of ab32503 staining Bax in rat kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32503, 1:250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Lane 1 = Bax protein (Tagged) (ab85157), 10 ng. Lane 2 = Extract of HeLa cells, 40 ug. Lane 3 = Extract of HepG2 cells, 40 ug. Lane 4 = Bax protein (Tagged) (ab85157), 10 ng. Lane 5 = Extract of HeLa cells, 40 ug. Lane 6 = Extract of HepG2 cells, 40 ug. SDS PAGE performed under reducing conditions (100mM DTT Sample heated at 50oC). Primary : Lanes 1-3: Anti Bax antibody (ab77566) at 1 ug/mL. Lanes 4-6: Anti Bax antibody (ab32503) at 1:2000 dilution. Secondary : Lanes 1-3: Goat anti mouse IgG(H&L)-HRP at 1:10000. Lanes 4-6: Goat anti rabbit IgG(H&L)-HRP at 1:10000. Development: ECL with 2 min exposure. Blocking: in 5% Milk + PBS for 3 hours at RT. Primary antibody: in 5% Milk + PBS overnight at 4 C. Secondary antibody: in 5% Milk + PBS for 2 hour at RT. Predicted band size : Bax 21kDa and Bax (Tagged) 49 kDa. Observed band size : Bax 21kDa and Bax (Tagged) 49 kDa.
All lanes : Anti-Bax antibody [E63] (ab32503) at 1/2000 dilution (purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell). Whole cell lysates
Lane 2 : Hep G2 (Human hepatocellular carcinoma epithelial cell). Whole cell lysates
Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) Whole cell lysates
Lane 4 : A549 (Human lung carcinoma epithelial cell) Whole cell lysates
Lane 5 : C2C12 (Mouse myoblasts myoblast) Whole cell lysates
Lane 6 : C6 (Rat glial tumor glial cell) Whole cell lysates
Lane 7 : Mouse brain. Whole tissue lysate
Lanes 8 & 10 : Rat brain. Whole tissue lysate
Lanes 9 & 11 : Rat Spleen. Whole tissue lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 21 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
Blocking and Diluting buffers: 5% NFDM/TBST
Exposure time 1~9 lanes 32 s; 10~11 lanes 3 min
Jurkat is negative reported by PMID: 15528359. Brain is low expressed reported by PMID: 27069530.
Immunohistochemical analysis of paraffin-embedded human lymph node using anti-Bax Rabbit Monoclonal Antibody (ab32503) at 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Standard Curve for Bax (Analyte: ab85157) dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [2D2] to Bax - BSA and Azide free (ab77566) at 0.2ug/ml and Detector Antibody Rabbit monoclonal [E63] to Bax (ab32503) at 0.5ug/ml Concentration of ab32503 may vary from lot to lot; please use this curve as guideline.
This product has been referenced in:
- Tian Y et al. Silencing of RHEB inhibits cell proliferation and promotes apoptosis in colorectal cancer cells via inhibition of the mTOR signaling pathway. J Cell Physiol 235:442-453 (2020). Read more (PubMed: 31332784) »
- Li Y et al. LncRNA-LET relieves hypoxia-induced injury in H9c2 cells through regulation of miR-138. J Cell Biochem 121:259-268 (2020). Read more (PubMed: 31222827) »