Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Bcl-2 antibody [EPR17509] (HRP) (ab209039)

Overview

  • Product name

    Anti-Bcl-2 antibody [EPR17509] (HRP)
    See all Bcl-2 primary antibodies
  • Description

    Rabbit monoclonal [EPR17509] to Bcl-2 (HRP)
  • Host species

    Rabbit
  • Conjugation

    HRP
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment within Human Bcl-2 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P10415

  • Positive control

    • WB: THP1 whole cell and Human Tonsil, Human Thymus and Mouse Spleen tissue lysates. IHC-P: normal human spleen tissue sections
  • General notes

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

Applications

Our Abpromise guarantee covers the use of ab209039 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
WB 1/5000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release (PubMed:17418785).
  • Tissue specificity

    Expressed in a variety of tissues.
  • Involvement in disease

    A chromosomal aberration involving BCL2 has been found in chronic lymphatic leukemia. Translocation t(14;18)(q32;q21) with immunoglobulin gene regions. BCL2 mutations found in non-Hodgkin lymphomas carrying the chromosomal translocation could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions.
  • Sequence similarities

    Belongs to the Bcl-2 family.
  • Domain

    BH1 and BH2 domains are required for the interaction with BAX and for anti-apoptotic activity.
    The BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3.
    The loop between motifs BH4 and BH3 is required for the interaction with NLRP1.
  • Post-translational
    modifications

    Phosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. Dephosphorylated by protein phosphatase 2A (PP2A).
    Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity.
    Monoubiquitinated by PARK2, leading to increase its stability. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome.
  • Cellular localization

    Mitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Apoptosis regulator Bcl 2 antibody
    • Apoptosis regulator Bcl-2 antibody
    • Apoptosis regulator Bcl2 antibody
    • AW986256 antibody
    • B cell CLL/lymphoma 2 antibody
    • B cell leukemia/lymphoma 2 antibody
    • Bcl-2 antibody
    • Bcl2 antibody
    • BCL2_HUMAN antibody
    • C430015F12Rik antibody
    • D630044D05Rik antibody
    • D830018M01Rik antibody
    • Leukemia/lymphoma, B-cell, 2 antibody
    • Oncogene B-cell leukemia 2 antibody
    • PPP1R50 antibody
    • Protein phosphatase 1 regulatory subunit 50 antibody
    • Protein phosphatase 1, regulatory subunit 50 antibody
    see all

Images

  • All lanes : Anti-Bcl-2 antibody [EPR17509] (HRP) (ab209039) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : BCL2 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 26 kDa


    Exposure time: 20 minutes


    ab209039 was shown to specifically react with Bcl-2 in wild-type HAP1 cells as signal was lost in BCL2 knockout cells. Wile-type and BCL2 knockout samples were subjected to SDS-PAGE. Ab209039 was incubated overnight at 4°C at 1/5000 dilution. Blots were developed with ECL technique.

  • IHC image of Bcl-2 staining in a section of formalin-fixed paraffin-embedded normal human spleen*, performed on a Leica BOND™. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH9, epitope retrieval solution 2) for 20mins. The section was then incubated with ab209039, 1/250 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    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.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-Bcl-2 antibody [EPR17509] (HRP) (ab209039) at 1/5000 dilution

    Lane 1 : THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate
    Lane 2 : Tonsil (Human) Tissue Lysate - adult normal tissue
    Lane 3 : Thymus (Human) Tissue Lysate - adult normal tissue
    Lane 4 : Spleen (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 26 kDa
    Observed band size: 26 kDa


    Exposure time: 8 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab209039 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

ab209039 has not yet been referenced specifically in any publications.

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