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  1. Link

    bcl-2-antibody-nor-235j-ab241548.pdf

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Cell Biology Apoptosis Intracellular Bcl2 Family
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Validated using a knockout cell lineRecombinant

Recombinant Anti-Bcl-2 antibody [NOR 235J] (ab241548)

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Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)
  • Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [NOR 235J] (ab241548)
  • Anti-Bcl-2 antibody [NOR 235J] (ab241548)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rat monoclonal [NOR 235J] to Bcl-2
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

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Overview

  • Product name

    Anti-Bcl-2 antibody [NOR 235J]
    See all Bcl-2 primary antibodies
  • Description

    Rat monoclonal [NOR 235J] to Bcl-2
  • Host species

    Rat
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein. This information is considered to be commercially sensitive.

  • Positive control

    • IHC-P: Human tonsil tissue. WB: Human spleen tissue, THP-1 cell line.
  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    NOR 235J
  • Isotype

    IgG2b
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Bcl2 Family
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Bcl 2 family
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Cell survival & death
    • Cancer
    • Tumor biomarkers
    • Oncoproteins
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Cancer
    • Cancer Metabolism
    • Cellular metabolic process
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Apoptosis marker and proteins ELISA kits
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • 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

    • Anti-Bcl-2 antibody [NOR 235J] - BSA and Azide free (ab252802)
  • Related Products

    • Anti-Bcl-2 antibody [EPR17509] (ab182858)
    • Anti-Bcl-2 antibody [E17] (ab32124)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab241548 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 a concentration of 0.98 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 26 kDa.
Notes
IHC-P
Use a concentration of 0.98 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 26 kDa.

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.
  • Target information above from: UniProt accession P10415 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 596 Human
    • Omim: 151430 Human
    • SwissProt: P10415 Human
    • Unigene: 150749 Human
    • 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 antibody [NOR 235J] (ab241548)
      Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)
      All lanes : Anti-Bcl-2 antibody [NOR 235J] (ab241548) at 1/1000 dilution

      Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
      Lane 2 : U937 (human histiocytic lymphoma monocyte), Whole cell lysate
      Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
      Lane 4 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
      Lane 5 : Human tonsil tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution (Goat Anti-Rat IgG (H+L), HRP))

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



      Exposure time:

      Lane 1,2,5:3 seconds

      Lane 3,4:10 seconds

      Blocking buffer / Diluting buffer and concentration:

      5% NFDM/TBST

    • Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)
      Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)
      All lanes : Anti-Bcl-2 antibody [NOR 235J] (ab241548) at 0.5 µg/ml

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : BCL2 knockout HAP1 whole cell lysate
      Lane 3 : NIH/3T3 whole cell lysate
      Lane 4 : THP-1 whole cell lysate
      Lane 5 : Human Spleen tissue lysate
      Lane 6 : Mouse Spleen tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rat IgG H&L (IRDye® 800CW ) (ab253031) at 1/20000 dilution

      Predicted band size: 26 kDa



      Lanes 1 - 6: Merged signal (red and green). Green - ab241548 observed at 26 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

      ab241548 was shown to react with Bcl-2 in Western blot. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween) before incubation with ab241548 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 0.5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat Anti-Rat IgG H&L (IRDye 800CW) (ab253031) and Goat anti-Rabbit IgG H&L (IRDye 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [NOR 235J] (ab241548)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [NOR 235J] (ab241548)

      IHC image of Bcl-2 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BOND™ system using the standard Protocol F.

      The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab241548, 0.98 µg/ml, 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 hematoxylin and mounted with DPX.

      The inset secondary-only 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.

    • Anti-Bcl-2 antibody [NOR 235J] (ab241548)
      Anti-Bcl-2 antibody [NOR 235J] (ab241548)

    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

    Datasheets and documents

    • Datasheet download

      Download

    References (0)

    Publishing research using ab241548? Please let us know so that we can cite the reference in this datasheet.

    ab241548 has not yet been referenced specifically in any publications.

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