RecombinantRabMAb

Recombinant Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)

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Western blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
  • Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
  • Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
  • Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
  • Immunoprecipitation - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
  • Dot Blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
  • Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21162] to Bcl-2 (phospho S70)
  • Suitable for: Flow Cyt (Intra), WB, Dot blot, ICC/IF, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Bcl-2 (phospho S70) antibody [EPR21162]
    See all Bcl-2 primary antibodies

  • Description

    Rabbit monoclonal [EPR21162] to Bcl-2 (phospho S70)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, Dot blot, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: Paclitaxel treated Jurkat whole cell lysate. Dot: Bcl-2 (phospho S70) peptide. ICC/IF: HeLa cells. Flow: HeLa cells. IP: Paclitaxel treated Jurkat whole cell lysate.

  • General notes

    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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab218123 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
Flow Cyt (Intra)
1/500.
WB
1/1000. Predicted molecular weight: 26 kDa.
Dot blot
1/1000.
ICC/IF
1/1000.
IP
1/30.
Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Predicted molecular weight: 26 kDa.
Dot blot
1/1000.
ICC/IF
1/1000.
IP
1/30.

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

    • Western blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
      Western blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
      All lanes : Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123) at 1/5000 dilution

      Lane 1 : Untreated Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
      Lane 2 : Jurkat treated with 1 µM paclitaxel for 24 hours whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 26 kDa


      Exposure time: 32 seconds


      Blocking/Dilution buffer and concentration: 2% BSA/TBST

      The expression profile observed is consistent with what has been described in the literature (PMID: 10097113).

    • Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
      Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (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.

    • Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
      Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)

      Intracellular 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) 

       

       

    • Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
      Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)

      Intracellular 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). 

       

       

    • Immunoprecipitation - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
      Immunoprecipitation - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (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.

    • Dot Blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
      Dot Blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (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.

    • Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
      Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (1)

    Publishing research using ab218123? Please let us know so that we can cite the reference in this datasheet.

    ab218123 has been referenced in 1 publication.

    • Chu X  et al. GSK-J4 induces cell cycle arrest and apoptosis via ER stress and the synergism between GSK-J4 and decitabine in acute myeloid leukemia KG-1a cells. Cancer Cell Int 20:209 (2020). PubMed: 32514253

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