The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/500.
1/500 - 1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
1/5000. PubMed: 22629415
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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.
Ab53154 staining human normal gallbladder tissue. Staining is localised to cell membrane and cytoplasm. Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be requir
ELISA - Anti-Bax antibody (ab53154)Image from Adams SM et al., PLoS One. 2012;7(5):e37540. Epub 2012 May 22. Fig 3.; doi:10.1371/journal.pone.0037540; May 22, 2012, PLoS ONE 7(5): e37540.
ELISA analysis of primary cultured rat cortical neurons, detecting Bax using ab53154. Cells were treated with (Tat) or without Tat (control) and either Estradiol, Genistein or Daidzein. Lysates were prepared after 24 hours. A 96-well plate was coated using carbonate coating buffer. 20 µg cell lysate was added to wells and incubated overnight at 4°C. Plates were blocked with 1% BSA for 2 hr at room temperature. Primary antibody (1/5000) was added to sample wells before incubating overnight at 4°C. A goat anti-rabbit ALP-conjugated IgG was used as secondary antibody and phosphatase substrate mixture was added. Absorbance was read at 650nm.
References for Anti-Bax antibody (ab53154)
This product has been referenced in:
Adams SM et al. Soy isoflavones genistein and daidzein exert anti-apoptotic actions via a selective ER-mediated mechanism in neurons following HIV-1 Tat(1-86) exposure. PLoS One7:e37540 (2012).
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