Recombinant
RabMAb

Recombinant Anti-Bcl-2 (phospho S70) antibody [EPR21162] - BSA and Azide free (ab233694)

Overview

  • Product name

    Anti-Bcl-2 (phospho S70) antibody [EPR21162] - BSA and Azide free
    See all Bcl-2 primary antibodies
  • Description

    Rabbit monoclonal [EPR21162] to Bcl-2 (phospho S70) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Dot blot, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Bcl-2 aa 50-150. The exact sequence is proprietary.
    Database link: P10415

  • Positive control

    • ICC/IF: HeLa cells.
  • General notes

    Ab233694 is the carrier-free version of ab218123. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab233694 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab233694 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 Use at an assay dependent concentration. Predicted molecular weight: 26 kDa.
Dot blot Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

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

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte) treated with 1 μM paclitaxel for 24h (Right) / Untreated control (Left) cell line labeling Bcl-2 (phospho S70) with ab218123 at 1/500 dilution. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    Square gate shows Bcl-2 (phospho S70) positive signal. The expression profile is consistent with what has been described in the literature (PMID: 10097113)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218123).

  • Dot blot analysis of Bcl-2 (phospho S70) labeled with ab218123 at 1/1000 dilution.

    Lane 1: Bcl-2 (phospho S70) peptide.
    Lane 2: Bcl-2 non-phospho peptide.

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100,000 dilution was used as secondary antibody.

    Blocking and dilution buffer: 2% BSA/TBST.

    Exposure time: 58 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218123).

  • Bcl-2 (phospho S70) was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cells from peripheral blood) treated with 1 μM paclitaxel for 24 hours whole cell lysate with ab218123 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab218123 at 0.5 μg/ml. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: Jurkat treated with 1 μM paclitaxel for 24 hours whole cell lysate 10 µg (Input). 

    Lane 2: ab218123 IP in Jurkat treated with 1 μM paclitaxel for 24 hours whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab218123 in Jurkat treated with 1 μM paclitaxel for 24 hours whole cell lysate.

    Blocking/ Dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 15 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218123).

  • Flow cytometric analysis of 80% methanol-fixed, 0.1% Tween-20 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Bcl-2 (phospho S70) with ab218123 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    Cells were pretreated with 20 μg/ml RNase A for 30 minutes to eliminate the non-specific binding between RNA and propidium iodide (PI).

    Bcl-2 (phospho S70) is highly expressed in mitotic cells (PMID: 10567572).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218123).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Bcl-2 (phospho S70) with ab218123 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/100 dilution (green). Confocal image showing positive staining in HeLa cells in M phase. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab195889) at 1/200 dilution (red). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab218123).

References

ab233694 has not yet been referenced specifically in any publications.

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