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    caspase-3-antibody-ab13847.pdf

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Validated using a knockout cell line

Anti-Caspase-3 antibody (ab13847)

  • Datasheet
  • SDS
Reviews (54)Q&A (2)References (966)

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Western blot - Anti-Caspase-3 antibody (ab13847)
  • Western blot - Anti-Caspase-3 antibody (ab13847)

Key features and details

  • Rabbit polyclonal to Caspase-3
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Caspase-3 antibody [E87] (ab32351)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Caspase-3 antibody
    See all Caspase-3 primary antibodies
  • Description

    Rabbit polyclonal to Caspase-3
  • Host species

    Rabbit
  • Specificity

    Stimulation is required to allow detection of the 17kDa cleaved form of the protein.  Please see images below for recommended treatment conditions and positive controls.

    ab13847 recognizes a cleaved form of Caspase 3 (~17 kDa) after apoptosis has been induced in wildtype cells and not Caspase 3 knockout cells.

    Some customers have used this antibody successfully in IHC-P however our latest tests were unsuccessful and therefore we can no longer guarantee this application. We would recommend ab32351 and ab184787 as an alternative product for this application.

  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Dog, Pig, Xenopus laevis, Drosophila melanogaster, Indian muntjac, Zebrafish, Rhesus monkey, Chinese hamster, Common marmoset, Schmidtea mediterranea, Salvelinus alpinus
  • Immunogen

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

  • Positive control

    • WB: Hap-1 WT, Human Caspase 3 (active) Recombinant Protein ICC: HeLa cells treated with 1 mM staurosporine
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Caspases
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Caspases
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Other proteases
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Caspases
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Signal Transduction
    • Antibodies
    • caspase-3
    • Neuroscience
    • Processes

Associated products

  • Assay kits

    • Caspase-3 Assay Kit (Colorimetric) (ab39401)
    • Cleaved Caspase-3 Staining Kit (FITC) (ab65613)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Caspase-3 protein (BSA and azide free) (denatured) (ab180336)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Quercetin, Antioxidative flavonoid (ab120247)
    • RTIL™ 13, dynamin I and II inhibitor (ab120465)
    • Z-IETD-FMK, Caspase-8 inhibitor (ab141382)
    • Z-VDVAD-FMK, caspase-2 inhibitor (ab142035)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab13847 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 (15)
1/500. Detects a band of approximately 17, 34 kDa (predicted molecular weight: 17, 34 kDa).
Notes
WB
1/500. Detects a band of approximately 17, 34 kDa (predicted molecular weight: 17, 34 kDa).

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.
  • 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.
  • Target information above from: UniProt accession P42574 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 836 Human
    • Entrez Gene: 12367 Mouse
    • Entrez Gene: 397244 Pig
    • Entrez Gene: 25402 Rat
    • Entrez Gene: 397720 Xenopus laevis
    • Entrez Gene: 140621 Zebrafish
    • Omim: 600636 Human
    • SwissProt: P42574 Human
    • SwissProt: P70677 Mouse
    • SwissProt: Q95ND5 Pig
    • SwissProt: P55213 Rat
    • SwissProt: P55866 Xenopus laevis
    • Unigene: 141125 Human
    • Unigene: 34405 Mouse
    • Unigene: 10562 Rat
    • Unigene: 17957 Xenopus laevis
    see all
  • Alternative names

    • A830040C14Rik antibody
    • Apopain antibody
    • CASP 3 antibody
    • CASP-3 antibody
    • CASP3 antibody
    • CASP3_HUMAN antibody
    • Casp3a antibody
    • Caspase 3 antibody
    • Caspase 3, apoptosis-related cysteine peptidase antibody
    • Caspase 3, apoptosis-related cysteine protease antibody
    • Caspase 3, apoptosis-related cysteine protease a antibody
    • Caspase-3 subunit p12 antibody
    • Caspase3 antibody
    • CC3 antibody
    • CPP 32 antibody
    • CPP-32 antibody
    • CPP32 antibody
    • CPP32B antibody
    • Cysteine protease CPP32 antibody
    • EC 3.4.22.56 antibody
    • ICE3 antibody
    • LICE antibody
    • mldy antibody
    • OTTHUMP00000165052 antibody
    • OTTHUMP00000165053 antibody
    • OTTHUMP00000165054 antibody
    • PARP cleavage protease antibody
    • Procaspase3 antibody
    • Protein Yama antibody
    • SCA 1 antibody
    • SCA-1 antibody
    • SCA1 antibody
    • SREBP cleavage activity 1 antibody
    • Yama antibody
    • Yama protein antibody
    see all

Images

  • Western blot - Anti-Caspase-3 antibody (ab13847)
    Western blot - Anti-Caspase-3 antibody (ab13847)

    Lane 1: Wild-type HAP1 cell lysate + Staurosporine (ab146588)  (1μM for 4h)
    Lane 2: Wild-type HAP1 cell lysate
    Lane 3: Caspase-3 knockout HAP1 cell lysate + Staurosporine (ab146588) (1μM for 4h)
    Lane 4: Caspase-3 knockout HAP1 cell lysate

    Lanes 1 - 4: Merged signal (red and green). Green - ab13847 observed at 17 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab13847 was shown to recognise Caspase 3 when Caspase 3 knockout samples were used, along with additional cross-reactive bands. Wild-type and Caspase 3 knockout samples (±staurosporine treatment) were subjected to SDS-PAGE. ab13847 and ab8245 (loading control to GAPDH) were diluted to 1/500 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Caspase-3 antibody (ab13847)
    Western blot - Anti-Caspase-3 antibody (ab13847)
    All lanes : Anti-Caspase-3 antibody (ab13847) at 1 µg/ml

    Lane 1 : Human Caspase 3 (active) Recombinant Protein
    Lane 2 : Human Pro Caspase 3 (inactive) Recombinant Protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 17, 34 kDa
    Observed band size: 17,32 kDa why is the actual band size different from the predicted?


    Exposure time: 8 minutes


    Caspase 3 exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce large (17kDa) and small (12kDa) subunits. These subunits dimerize to form the active enzyme. ab13847 specifically detects the large active subunit (17kDa) and the  inactive pro Caspase 3 (32 kDa).

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab13847 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

     

    Secondary antibody - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (966)

Publishing research using ab13847? Please let us know so that we can cite the reference in this datasheet.

ab13847 has been referenced in 966 publications.

  • Liu Y  et al. Paeonol inhibits apoptosis of vascular smooth muscle cells via up-regulation of autophagy by activating class III PI3K/Beclin-1 signaling pathway. Life Sci 264:118714 (2021). PubMed: 33157088
  • Zhang T  et al. Polysaccharides extracted from Rheum tanguticum ameliorate radiation-induced enteritis via activation of Nrf2/HO-1. J Radiat Res 62:46-57 (2021). PubMed: 33140083
  • Gao W  et al. Procyanidin B1 promotes in vitro maturation of pig oocytes by reducing oxidative stress. Mol Reprod Dev 88:55-66 (2021). PubMed: 33241626
  • Jin P  et al. Salidroside inhibits apoptosis and autophagy of cardiomyocyte by regulation of circular RNA hsa_circ_0000064 in cardiac ischemia-reperfusion injury. Gene 767:145075 (2021). PubMed: 32858179
  • Li J  et al. Overexpression of lncRNA Dancr inhibits apoptosis and enhances autophagy to protect cardiomyocytes from endoplasmic reticulum stress injury via sponging microRNA-6324. Mol Med Rep 23:N/A (2021). PubMed: 33300079
View all Publications for this product

Customer reviews and Q&As

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Submit a review Submit a question

1-10 of 56 Abreviews or Q&A

IHC - Wholemount abreview for Anti-active + pro Caspase 3 antibody

Good
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Salvelinus alpinus Embryo (Developing head at a pre-hatching embryonic stage)
Specification
Developing head at a pre-hatching embryonic stage
Read More

Mr. Ehsan Pashay Ahi

Verified customer

Submitted Jul 28 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Caspase-3 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (brain organoid)
Permeabilization
Yes - 0.3%tritonX-100
Specification
brain organoid
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jan 18 2022

Immunocytochemistry/ Immunofluorescence abreview for Anti-Caspase-3 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mice hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Mice hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More

Dr. Armen Petrosyan

Verified customer

Submitted Aug 03 2020

Western blot abreview for Anti-Caspase-3 antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Treatment
4mg/ml bevacizumab for 48hrs
Specification
Glioblastoma cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Jul 09 2020

Immunohistochemistry free floating abreview for Anti-Caspase-3 antibody

Average
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry free floating
Sample
Rat Tissue sections (Brain (Motorcortex) 40µm)
Specification
Brain (Motorcortex) 40µm
Read More

Abcam user community

Verified customer

Submitted May 28 2020

Western blot abreview for Anti-Caspase-3 antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Triple negative breast cancer)
Gel Running Conditions
Non-reduced Denaturing (15)
Loading amount
10 µg
Treatment
20µM Compound for 24hrs
Specification
Triple negative breast cancer
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Dr. yushan lin

Verified customer

Submitted Apr 20 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Caspase-3 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Kidney H9 Organoids)
Permeabilization
Yes - 0.05% Tween 20
Specification
Kidney H9 Organoids
Blocking step
Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Feb 26 2020

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Caspase-3 antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (brain)
Antigen retrieval step
None
Permeabilization
Yes - triton 1%
Specification
brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5 · Temperature: 20°C
Fixative
Paraformaldehyde
Read More

MRS. Francoise Geffroy

Verified customer

Submitted Feb 19 2020

Flow Cytometry abreview for Anti-Caspase-3 antibody

Good
Abreviews
Abreviews
abreview image
Application
Flow Cytometry
Sample
Human Cell (PBMCs)
Permeabilization
Yes - perm buffer
Gating Strategy
Gate on Lymphozytes and monocytes
Specification
PBMCs
Preparation
Cell harvesting/tissue preparation method: Ficoll density separation
Sample buffer: PBS
Fixation
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jan 25 2020

Immunohistochemistry (Frozen sections) abreview for Anti-Caspase-3 antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin biopsy)
Permeabilization
Yes - 0,1% Triton
Specification
Skin biopsy
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jan 21 2020

1-10 of 56 Abreviews or Q&A

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