Recombinant
RabMAb

Recombinant Anti-Caspase-3 p12 antibody [EPR16888] (ab179517)

Overview

  • Product name

    Anti-Caspase-3 p12 antibody [EPR16888]
    See all Caspase-3 p12 primary antibodies
  • Description

    Rabbit monoclonal [EPR16888] to Caspase-3 p12
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human Caspase-3 p12 aa 150 to the C-terminus. The exact sequence is proprietary.
    Database link: P42574

  • Positive control

    • WB: HeLa and NIH/3T3 whole cell lysates. Mouse brain, heart and spleen; and Rat brain and heart lysates. IHC-P: Human tonsil, mouse testis and rat kidney tissues. ICC/IF: K562 cells.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR16888
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab179517 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 1/1000. Detects a band of approximately 32 ,29, 12 kDa (predicted molecular weight: 32, 29, 12 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/150.
Flow Cyt Use at an assay dependent concentration.

Target

  • Function

    Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-
    -Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. Triggers cell adhesion in sympathetic neurons through RET cleavage.
  • Tissue specificity

    Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.
  • Sequence similarities

    Belongs to the peptidase C14A family.
  • Post-translational
    modifications

    Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
    S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Form

    Caspase-3 p12 is from aa 176-277 of P42574.
  • Alternative names

    • Apopain antibody
    • CASP-3 antibody
    • CASP3 antibody
    • CASP3_HUMAN antibody
    • Caspase 3 p12 antibody
    • Caspase-3 subunit p12 antibody
    • CPP-32 antibody
    • Cysteine protease CPP32 antibody
    • Protein Yama antibody
    • SCA-1 antibody
    • SREBP cleavage activity 1 antibody
    see all

Images

  • All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates treated with staurosporine 1uM for 4hr
    Lane 2 : HeLa untreated whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 32, 29, 12 kDa
    Observed band size: 12,29,32 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

     

    32 and 29 kDa bands represent procaspase-3; 12-kDa band is Caspase-3 subunit p12.

  • Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) with purified ab179517 at 1/240 dilution (Red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Caspase-3 p12 with ab179517 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing cytoplasmic staining on K562 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    -ve control 1: ab179517 at 1/150 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

     

  • All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution

    Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) treated with staurosporine 1uM for 4h
    Lane 2 : Untreated NIH/3T3 whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 32, 29, 12 kDa
    Observed band size: 12,29,32 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

     

    32 and 29 kDa bands represent procaspase-3; 12-kDa band is Caspase-3 subunit p12.

  • Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/5000 dilution + Mouse spleen lysates at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 32, 29, 12 kDa
    Observed band size: 29,32 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

     

    32 and 29 kDa bands represent procaspase-3.

  • All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution

    Lane 1 : Mouse brain lysates
    Lane 2 : Mouse heart lysates
    Lane 3 : Rat brain lysates
    Lane 4 : Rat heart lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 32, 29, 12 kDa
    Observed band size: 29,32 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

     

    32 and 29 kDa bands represent procaspase-3.

  • Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining in germinal center, weaker staining in the mantle zone (MZ) lymphocytes is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on leydig cells and spermatogonial cells is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Granular staining on epithelium from kidney is observed. Counter stained with Hematoxylin.

    Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution.

     

    [1] Afshin Samali, Boris Zhivotovsky, Dean P. Jonesa, Sten Orreniusa. Detection of pro-caspase-3 in cytosol and mitochondria of various tissues (rat). FEBS Letters 431 (1998) 167-169.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Gao X  et al. Aconitine induces apoptosis in H9c2 cardiac cells via mitochondria-mediated pathway. Mol Med Rep 17:284-292 (2018). Read more (PubMed: 29115599) »
  • Liu J  et al. BECN1-dependent CASP2 incomplete autophagy induction by binding to rabies virus phosphoprotein. Autophagy 13:739-753 (2017). Read more (PubMed: 28129024) »
See all 4 Publications for this product

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