Anti-active Caspase-3 antibody [E83-77] (ab32042)

Overview

  • Product nameAnti-active Caspase-3 antibody [E83-77]
    See all active Caspase-3 primary antibodies
  • Description
    Rabbit monoclonal [E83-77] to active Caspase-3
  • SpecificityMouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information This antibody only detects the active (cleaved) form of Caspase-3 and does not recognize the pro form of Caspase-3. PubMedID 19789217 describes the detection of human cells injected into mice.
  • Tested applicationsSuitable for: IHC-Fr, WB, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human active Caspase-3 aa 1-100 (N terminal). A synthetic peptide corresponding to residues following Ser29 of human Caspase 3 (N terminus of p17 subunit).
    Database link: P42574

  • Positive control
    • WB: Jurkat cell lysate (camptothecin treated); HeLa cell lysate (staurosporine treated). IF: Hela cells (staurosporine treated).
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab32042 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-Fr Use at an assay dependent concentration.
WB 1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 32 kDa).
ICC/IF 1/100 - 1/250.
  • Application notesIs unsuitable for Flow Cyt or IP.
  • Target

    • FunctionInvolved 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 specificityHighly 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 similaritiesBelongs 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 localizationCytoplasm.
    • Information by UniProt
    • Database links
    • Alternative names
      • Apopain antibody
      • CASP 3 antibody
      • CASP-3 antibody
      • CASP3 antibody
      • CASP3_HUMAN antibody
      • Caspase 3 antibody
      • Caspase-3 subunit p12 antibody
      • CPP 32 antibody
      • CPP-32 antibody
      • CPP32B antibody
      • Cysteine protease CPP32 antibody
      • PARP cleavage protease antibody
      • Protein Yama antibody
      • SCA-1 antibody
      • SCA1 antibody
      • SREBP cleavage activity 1 antibody
      • Yama antibody
      see all

    Anti-active Caspase-3 antibody [E83-77] images

    • Ab32042, at dilution of 1/100, staining HeLa cells by Immunofluorescence.
      Left image: control.
      Right image: staurosporine treated.
    • All lanes : Anti-active Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution

      Lane 1 : HeLa Whole Cell Lysate (2 uM Staurosporine, 4Hr) at 20 µg
      Lane 2 : HeLa Whole Cell Lysate (untreated) at 20 µg
      Lane 3 : Active Caspase 3 (recombinant protein) at 0.1 µg

      Secondary
      800CW Goat Anti-Rabbit IgG at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size : 32 kDa
      Additional bands at : 17 kDa (possible mature (processed) protein).
    • Lanes 1 - 2 : anti Pro Caspase 3 at 1/10000 dilution
      Lanes 3 - 4 : Anti-active Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : Jurkat cell lysate + Camptothecin
      Lane 3 : Jurkat cell lysate
      Lane 4 : Jurkat cell lysate + Camptothecin


      Predicted band size : 32 kDa
      Observed band size : 17 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 30 kDa. We are unsure as to the identity of these extra bands.

    Protocols

    References for Anti-active Caspase-3 antibody [E83-77] (ab32042)

    This product has been referenced in:
    • Wang X  et al. Therapeutic Potential of Delivering Arsenic Trioxide into HPV-Infected Cervical Cancer Cells Using Liposomal Nanotechnology. Nanoscale Res Lett 11:94 (2016). ICC/IF ; Human . Read more (PubMed: 26887578) »
    • Tura A  et al. Biocompatibility of the vital dye Acid Violet-17 on retinal pigment epithelial cells. Clin Ophthalmol 10:1435-45 (2016). Read more (PubMed: 27536056) »

    See all 22 Publications for this product

    Product Wall

    Thank you for your enquiry.

    I would not recommend using an autoclave, as this could damage the tissue. I would suggest using a pressure cooker or a microwave. The time and temperature may require some optimization. For the time, I can suggest...

    Read More
    Application Immunocytochemistry/ Immunofluorescence
    Sample Monkey Cell (Kidney)
    Permeabilization Yes - 0.1% triton x-100
    Specification Kidney
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 29 2016

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Western blot
    Sample Monkey Cell lysate - whole cell (Kidney)
    Gel Running Conditions Non-reduced Non-Denaturing (Native)
    Loading amount 30 µg
    Specification Kidney
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted May 06 2016

    Application Immunohistochemistry (Frozen sections)
    Sample Mouse Tissue sections (Brain)
    Permeabilization Yes - 0.3% Triton x 100: 30 min
    Specification Brain
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted May 06 2016

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (prostate cancer)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 0.05% trysin
    Permeabilization Yes - 0.1% Triton X-100
    Specification prostate cancer
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Dec 24 2015

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Rat Tissue sections (Thymus)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization No
    Specification Thymus
    Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative Formaldehyde
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Dec 08 2015

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (Human melanoma xenograft)
    Antigen retrieval step Heat mediated
    Permeabilization No
    Specification Human melanoma xenograft
    Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
    Fixative Formaldehyde
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Dec 08 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (Liver)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA pH9
    Permeabilization No
    Specification Liver
    Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Aug 13 2015

    Pro-caspase 3 is cleaved between amino acids 175 and 176 of the sequence described in the UniProt entry, P42574. http://www.uniprot.org/uniprot/P42574
    Antibody ab32042 is raised against the N terminus of subunit p17.

    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing
    Sample Human Cell lysate - whole cell (HT29)
    Specification HT29
    Treatment IGF-1R inhibitor
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Mar 21 2014

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