Overview

  • 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
    Does not react with: Mouse
  • 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.

  • 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.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    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
    10X Human Bax Capture Antibody 1 x 600µl
    10X Human Bax Detector Antibody 1 x 600µl
    Human Bax Lyophilized Recombinant Protein 1 x 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 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
  • 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
  • Alternative names
    • Apoptosis regulator BAX
    • BAX
    • Bax-protein
    • BAX_HUMAN
    • BAXA
    • Baxdelta2G9
    • Baxdelta2G9omega
    • Baxdelta2omega
    • Bcl-2-like protein 4
    • BCL2 associated X protein
    • BCL2 associated X protein omega
    • BCL2 associated X protein transcript variant delta2
    • Bcl2-L-4
    • BCL2L4
    • membrane isoform alpha
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab199080 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • Interpolated 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.

  • Cells 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.

Protocols

References

ab199080 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab199080.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up