Validated using a knockout cell line
Recombinant
RabMAb

Anti-BAT3 antibody [EPR9223] (ab137076)

Overview

  • Product name
    Anti-BAT3 antibody [EPR9223]
    See all BAT3 primary antibodies
  • Description
    Rabbit monoclonal [EPR9223] to BAT3
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Human BAT3 (C terminal).

  • Positive control
    • HeLa, A431; human fetal brain lysates; Human kidney and testis tissue, mouse and rat brain tissue lysate.
  • General notes

    The mouse and rat recommendation. This antibody may not be suitable for IHC with mouse or rat samples. 

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab137076 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
ICC/IF 1/500.
WB 1/1000 - 1/10000. Predicted molecular weight: 119 kDa.
IHC-P 1/50 - 1/100.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/30.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/100 - 1/1000. 

Target

  • Function
    Chaperone that plays a key role in various processes such as apoptosis, insertion of tail-anchored (TA) membrane proteins to the endoplasmic reticulum membrane and regulation of chromatin. Acts in part by regulating stability of proteins and their degradation by the proteasome. Participates in endoplasmic reticulum stress-induced apoptosis via its interaction with AIFM1/AIF by regulating AIFM1/AIF stability and preventing its degradation. Also required during spermatogenesis for synaptonemal complex assembly via its interaction with HSPA2, by inhibiting polyubiquitination and subsequent proteasomal degradation of HSPA2. Required for selective ubiquitin-mediated degradation of defective nascent chain polypeptides by the proteasome. In this context, may play a role in immuno-proteasomes to generate antigenic peptides via targeted degradation, thereby playing a role in antigen presentation in immune response. Key component of the BAG6/BAT3 complex, a cytosolic multiprotein complex involved in the post-translational delivery of tail-anchored (TA) membrane proteins to the endoplasmic reticulum membrane. TA membrane proteins, also named type II transmembrane proteins, contain a single C-terminal transmembrane region. BAG6/BAT3 acts by facilitating TA membrane proteins capture by ASNA1/TRC40: it is recruited to ribosomes synthesizing membrane proteins, interacts with the transmembrane region of newly released TA proteins and transfers them to ASNA1/TRC40 for targeting to the endoplasmic reticulum membrane. Also involved in DNA damage-induced apoptosis: following DNA damage, accumulates in the nucleus and forms a complex with p300/EP300, enhancing p300/EP300-mediated p53/TP53 acetylation leading to increase p53/TP53 transcriptional activity. When nuclear, may also act as a component of some chromatin regulator complex that regulates histone 3 'Lys-4' dimethylation (H3K4me2).
  • Sequence similarities
    Contains 1 ubiquitin-like domain.
  • Post-translational
    modifications
    Cleavage by caspase-3 releases a C-terminal peptide that plays a role in ricin-induced apoptosis.
    In case of infection by L.pneumophila, ubiquitinated by the SCF(LegU1) complex.
  • Cellular localization
    Cytoplasm > cytosol. Nucleus. The C-terminal fragment generated by caspase-3 is cytoplasmic.
  • Information by UniProt
  • Database links
  • Alternative names
    • 2410045D21Rik antibody
    • AA408914 antibody
    • BAG 6 antibody
    • BAG family molecular chaperone regulator 6 antibody
    • BAG-6 antibody
    • BAG6 antibody
    • BAG6_HUMAN antibody
    • BAT 3 antibody
    • BCL2-associated athanogene 6 antibody
    • D17H6S52E antibody
    • D6S52E antibody
    • G3 antibody
    • HLA B associated transcript 3 antibody
    • HLA-B associated transcript 3 antibody
    • HLA-B associated transcript-3 antibody
    • HLA-B-associated transcript 3 antibody
    • large proline rich protein BAG6 antibody
    • Large proline rich protein BAT3 antibody
    • Large proline-rich protein BAG6 antibody
    • large proline-rich protein BAT3 antibody
    • Protein G3 antibody
    • Protein Scythe antibody
    • Scythe antibody
    • Scythe, homolog of Xenopus antibody
    see all

Images



  • Predicted band size : 119 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: BAT3 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab137076 observed at 155 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab137076 was shown to specifically react with BAT3 when BAT3 knockout samples were used. Wild-type and BAT3 knockout samples were subjected to SDS-PAGE.  Ab137076 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of Paraffin-embedded human testis tissue sections labeling BAT3 with purified ab137076 at a dilution of 1/50 dilution (7.1 μg/ml). ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 was used as the secondary anitbody. Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated using EDTA Buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling BAT3 with purified ab137076 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200. DAPI (blue) was used as a nuclear counterstain. Secondary Only Control: PBS was used instead of the primary antibody as the negative control. 

  • Flow cytometry analysis of HeLa cells labelling BAT3 (red) with purified ab137076 at dilution of 1/30. Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 was used as the secondary antibody. Cells were fixed with 4% paraformaldehyde. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • Anti-BAT3 antibody [EPR9223] (ab137076) at 1/5000 dilution (purified) + Mouse brain tissue lysate at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 119 kDa
    Blocking/Diluting buffer 5% NFDM/TBST
  • All lanes : Anti-BAT3 antibody [EPR9223] (ab137076) at 1/1000 dilution (purified)

    Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
    Lane 2 : Human fetal brain tissue lysate
    Lane 3 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size : 119 kDa
    Observed band size : 160 kDa (why is the actual band size different from the predicted?)
    Blocking/Diluting buffer 5% NFDM/TBST
  • Overlay histogram showing Hela cells stained with ab137076 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab137076, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling BAT3 with unpurified ab137076 at 1/50 dilution.

  • All lanes : Anti-BAT3 antibody [EPR9223] (ab137076) at 1/1000 dilution (unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : Human fetal brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size : 119 kDa

References

ab137076 has not yet been referenced specifically in any publications.

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