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    bax-antibody-e63-bsa-and-azide-free-ab216985.pdf

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Cell Biology Apoptosis Intracellular Associated Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
  • Immunoprecipitation - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
  • Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
  • Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
  • Sandwich ELISA - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
  • Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E63] to Bax - BSA and Azide free
  • Suitable for: IHC-P, Sandwich ELISA, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

HRP Unconjugated

You may also be interested in

Protein
Product image
Recombinant Human Bax protein (ab173026)
Primary
Product image
Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
Conjugation
Product image
Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

View more associated products

Overview

  • Product name

    Anti-Bax antibody [E63] - BSA and Azide free
    See all Bax primary antibodies
  • Description

    Rabbit monoclonal [E63] to Bax - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Sandwich ELISA, IP, WBmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Recombinant Human Bax protein (Tagged) (ab85157), HeLa, HepG2, A549, C2C12 and C6 cell lysate. Wild-type Hap1 cell lysate. Rat spleen tissue lysate. IHC-P: Human lymph node and rat kidney tissues. Human lung carcinoma tissue. IP: HeLa cell lysate.
  • General notes

    ab216985 is the carrier-free version of ab32503.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E63
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Associated Proteins
    • Cell Biology
    • Apoptosis
    • Intracellular
    • Bcl2 Family
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Bcl 2 family
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Apoptosis marker and proteins ELISA kits
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Bcl 2 family
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Alternative Versions

    • HRP Anti-Bax antibody [E63] (ab205822)
    • Anti-Bax antibody [E63] (ab32503)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human BAX knockout HeLa cell line (ab255363)
  • KO cell lysates

    • Human BAX knockout HeLa cell lysate (ab263841)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
  • Recombinant Protein

    • Recombinant Human Bax protein (ab173026)
  • Related Products

    • 2-Methoxyantimycin A3, Bcl-xL and Bcl-2 inhibitor (ab141523)
    • Recombinant Human Bax protein (ab173026)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab216985 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
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Sandwich ELISA
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
Notes
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Sandwich ELISA
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

Target

  • 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.
  • Target information above from: UniProt accession Q07812 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 280730 Cow
    • Entrez Gene: 581 Human
    • Entrez Gene: 12028 Mouse
    • Entrez Gene: 24887 Rat
    • Omim: 600040 Human
    • SwissProt: O02703 Cow
    • SwissProt: Q07812 Human
    • SwissProt: Q07813 Mouse
    • SwissProt: Q63690 Rat
    • Unigene: 624291 Human
    • Unigene: 19904 Mouse
    • Unigene: 10668 Rat
    see all
  • Alternative names

    • Apoptosis regulator BAX antibody
    • BAX antibody
    • Bax-protein antibody
    • BAX_HUMAN antibody
    • BAXA antibody
    • Baxdelta2G9 antibody
    • Baxdelta2G9omega antibody
    • Baxdelta2omega antibody
    • Bcl-2-like protein 4 antibody
    • BCL2 associated X protein antibody
    • BCL2 associated X protein omega antibody
    • BCL2 associated X protein transcript variant delta2 antibody
    • Bcl2-L-4 antibody
    • BCL2L4 antibody
    • membrane isoform alpha antibody
    see all

Images

  • Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    All lanes : Anti-Bax antibody [E63] (ab32503) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : BAX knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 21 kDa
    Observed band size: 21 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab32503).

      Lanes 1- 2: Merged signal (red and green). Green - ab32503 observed at 21 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab32503 was shown to react with Bax in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255363 (knockout cell lysate ab263841) was used. Wild-type HeLa and BAX knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32503 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32503).

  • Immunoprecipitation - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Immunoprecipitation - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

    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 (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32503).

  • Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    All lanes : Anti-Bax antibody [E63] (ab32503) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : BAX knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 21 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab32503).

    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 in wild-type HAP1 cells, 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32503).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

    Immunohistochemical analysis of paraffin-embedded human lymph node using anti-Bax Rabbit Monoclonal Antibody (ab32503) at 1/250 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32503).

    Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Western blot - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    This data was developed using ab32503, the same antibody clone in a different buffer formulation. Different batches of ab32503 were tested on HeLa (Human cervix adenocarcinoma epithelial cell) lysate at 0.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 18 kDa.
  • Sandwich ELISA - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Sandwich ELISA - Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

    Standard Curve for Bax (Analyte: Recombinant human Bax protein (tagged) 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32503).

  • Anti-Bax antibody [E63] - BSA and Azide free (ab216985)
    Anti-Bax antibody [E63] - BSA and Azide free (ab216985)

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (1)

Publishing research using ab216985? Please let us know so that we can cite the reference in this datasheet.

ab216985 has been referenced in 1 publication.

  • Jiang X  et al. Chipmunk Brain Is Resistant to Injury from Deep Hypothermic Circulatory Arrest During Cardiopulmonary Bypass. Ther Hypothermia Temp Manag 9:118-127 (2019). PubMed: 30036167

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