Human Bax ELISA Kit (ab199080)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 2.2 pg/ml
- Range: 12.5 pg/ml - 800 pg/ml
- Sample type: Cell culture extracts, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human Bax ELISA Kit
See all Bax kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Tissue Ext. 3 3.8% Inter-assay Sample n Mean SD CV% Tissue Ext. 5 3.2% -
Sample type
Cell culture extracts, Tissue Extracts -
Assay type
Sandwich (quantitative) -
Sensitivity
2.2 pg/ml -
Range
12.5 pg/ml - 800 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture extracts 98 86% - 108% Tissue Extracts 99 87% - 109% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human Bax SimpleStep ELISA® kit (ab199080) has been re-developed with new capture and detector antibodies. We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantifying Bax protein in human cell and tissue extracts.
Bax in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Bax protein in human cell and tissue extracts.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Bax accelerates cell death by binding to and antagonizing the apoptosis repressor, BCL2. It undergoes a translocation to the mitochondrion membrane during cell stress, leading to the release of cytochrome c which then triggers apoptosis.
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Notes
Bax is a pro-apoptotic factor of Bcl-2 family. It accelerates programmed cell death by binding to, and antagonizing the apoptosis repressor BCL2 or its adenovirus homolog E1B 19k protein. Under stress conditions, Bax undergoes a conformation change that causes translocation to the mitochondrion membrane, leading to the release of cytochrome c that then triggers apoptosis. Bax promotes activation of CASP3, and thereby apoptosis. Under a normal conditions, Bax colocalizes with 14-3-3 proteins in the cytoplasm. Under stress conditions, Bax undergoes a conformation change that causes release from JNK-phosphorylated 14-3-3 proteins and translocation to the mitochondrion membrane. Bax forms higher oligomers under stress conditions. Bax interacts with BCL2L11. The interaction with BCL2L11 promotes BAX oligomerization and association with mitochondrial membranes, with subsequent release of cytochrome c. Bax forms heterodimers with BCL2, E1B 19K protein, BCL2L1 isoform Bcl-X(L), BCL2L2, MCL1 and A1. Bax interacts with SH3GLB1 and HN. Bax interacts with SFN and YWHAZ; the interaction occurs in the cytoplasm. Under stress conditions, JNK-mediated phosphorylation of SFN and YWHAZ, releases BAX to mitochondria. Bax isoform Sigma interacts with BCL2A1 and BCL2L1 isoform Bcl-X(L). Bax interacts with RNF144B, which regulates the ubiquitin-dependent stability of BAX. Bax interacts with CLU under stress conditions that cause a conformation change leading to BAX oligomerization and association with mitochondria. Bax does not interact with CLU in unstressed cells. Bax interacts with FAIM2/LFG2. Bax interacts with human cytomegalovirus/HHV-5 protein vMIA/UL37. Bax interacts with BOP/C22orf29.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human Bax Capture Antibody 1 x 600µl 10X Human Bax Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BI 1 x 6ml Human Bax Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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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. -
Domain
Intact 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 localization
Cytoplasm 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
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Alternative names
- Apoptosis regulator BAX
- BAX
- Bax-protein
see all -
Database links
- Entrez Gene: 581 Human
- Omim: 600040 Human
- SwissProt: Q07812 Human
- Unigene: 624291 Human
Images
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Other - Human Bax ELISA Kit (ab199080)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human Bax standard curve in 1X Cell Extraction Buffer PTRBackground-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Bax in human HepG2 cell extract, and HeLa cell extract and liver tissue extractInterpolated concentrations of native Bax in human HepG2 cell extract based on a 10 μg/mL extract load, and HeLa cell extract and liver tissue extract based on a 20 μg/mL extract load. The concentrations of Bax were measured in duplicate and interpolated from the Bax standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Bax concentration was determined to be 351.9 pg/mL in HepG2 cell extract, 358.5 pg/mL in HeLa cell extract, and 235.8 pg/mL in liver tissue extract.
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Comparison of Bax distribution in subcellular fractions derived from 3.7x103 HeLa cells and whole cells cultured in the presence (treated) or absence (untreated) of 1 µM staurosporine for 4 hoursCells were collected directly after treatment and subcellular fractions were prepared using a cell fractionation kit (ab109719). The concentrations of Bax were measured in three different dilutions of the fraction samples in duplicates and interpolated from the Bax standard curve. The interpolated values are plotted (mean +/- SD, n=3). The mean Bax concentration was determined be 79.66 pg/mL in the untreated cytosol fraction, 29.30 pg/mL in the untreated mitochondrial fraction, 152.1 pg/mL in the untreated whole cell sample, 23.15 pg/mL in the treated cytosol fraction, 631.5 pg/mL in the treated mitochondrial fraction, and 825.4 pg/mL in the treated whole cell sample.
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Sandwich ELISA - Human Bax ELISA Kit (ab199080)To learn more about the advantages of recombinant antibodies see here.
