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  1. Link

    caspase-7-antibody-e22-ab32522.pdf

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Cell Biology Apoptosis Intracellular Caspases etc Caspases
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Validated using a knockout cell lineRabMAb

Anti-Caspase-7 antibody [E22] (ab32522)

  • Datasheet
Reviews (2) Submit a question References (22)

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Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
  • Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
  • Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
  • Immunocytochemistry/ Immunofluorescence - Anti-Caspase-7 antibody [E22] (ab32522)
  • Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [E22] (ab32522)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [E22] (ab32522)
  • Immunoprecipitation - Anti-Caspase-7 antibody [E22] (ab32522)

Key features and details

  • Rabbit monoclonal [E22] to Caspase-7
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Caspase-7 antibody [E22]
    See all Caspase-7 primary antibodies
  • Description

    Rabbit monoclonal [E22] to Caspase-7
  • Host species

    Rabbit
  • Specificity

    The antibody should recognize both pro-form and p20 cleaved-form. The antibody does not cross-react with other Caspase family members.
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: Jurkat and HeLa whole cell lysate (ab150035). IHC-P: Human skin cancer tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: Jurkat cell lysates
  • 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.

    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

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E22
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Caspases
    • 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
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • HRP Anti-Caspase-7 antibody [E22] (ab206039)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CASP7 (Caspase-7) knockout HeLa cell line (ab265777)
  • KO cell lysates

    • Human CASP7 (Caspase-7) knockout HeLa cell lysate (ab257380)
  • Positive Controls

    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant human Caspase-7 protein (ab198478)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32522 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)
1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.
IP (1)
1/20.
Flow Cyt (Intra)
1/250.
Notes
WB
1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.
IP
1/20.
Flow Cyt (Intra)
1/250.

Target

  • Function

    Involved in the activation cascade of caspases responsible for apoptosis execution. Cleaves and activates sterol regulatory element binding proteins (SREBPs). Proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-
    -Gly-217' bond. Overexpression promotes programmed cell death.
  • Tissue specificity

    Highly expressed in lung, skeletal muscle, liver, kidney, spleen and heart, and moderately in testis. No expression in the brain.
  • Sequence similarities

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

    Cleavages by granzyme B or caspase-10 generate the two active subunits. Propeptide domains can also be cleaved efficiently by caspase-3. Active heterodimers between the small subunit of caspase-7 and the large subunit of caspase-3, and vice versa, also occur.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession P55210 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 840 Human
    • Omim: 601761 Human
    • SwissProt: P55210 Human
    • Unigene: 9216 Human
    • Alternative names

      • Apoptotic protease Mch-3 antibody
      • Apoptotic protease MCH3 antibody
      • CASP-7 antibody
      • CASP7 antibody
      • CASP7_HUMAN antibody
      • Caspase 7 antibody
      • Caspase 7 apoptosis related cysteine peptidase antibody
      • Caspase-7 subunit p11 antibody
      • Caspase7 antibody
      • CMH 1 antibody
      • CMH-1 antibody
      • CMH1 antibody
      • ICE LAP3 antibody
      • ICE-LAP3 antibody
      • ICE-like apoptotic protease 3 antibody
      • LICE2 antibody
      • MCH3 antibody
      see all

    Images

    • Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
      Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
      All lanes : Anti-Caspase-7 antibody [E22] (ab32522) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : CASP7 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 34 kDa
      Observed band size: 38 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab32522 observed at 38 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

       ab32522 was shown to react with pro Caspase-7 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265777 (knockout cell lysate ab257380) was used. Wild-type HeLa and CASP7 knockout HeLa cell lysates were subjected to SDS-PAGE. ab32522 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
      Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
      Anti-Caspase-7 antibody [E22] (ab32522) at 1/1000 dilution + Jurkat cell lysate

      Predicted band size: 34 kDa
      Observed band size: 34 kDa

    • Western blot - Anti-Caspase-7 antibody [E22] (ab32522)
      Western blot - Anti-Caspase-7 antibody [E22] (ab32522)

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Wild type HAP1 + Staurosporine ab120056 whole cell lysate (20 µg)
      Lane 3: CASP7 knockout HAP1 whole cell lysate (20 µg)
      Lane 4: CASP7 + Staurosporine knockout HAP1 whole cell lysate (20 µg)
      Lane 5: HeLa whole cell lysate (20 µg)
      Lane 6: HeLa + Staurosporine whole cell lysate (20 µg)

      Lanes 1 - 6: Merged signal (red and green). Green - ab32522 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab32522 was shown to specifically react with HAP1 + Staurosporine  when HAP1 + Staurosporine knockout samples were used. Wild-type and HAP1 + Staurosporine knockout samples were subjected to SDS-PAGE. Ab32522 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Caspase-7 antibody [E22] (ab32522)
      Immunocytochemistry/ Immunofluorescence - Anti-Caspase-7 antibody [E22] (ab32522)

      Immunofluorescent staining of HeLa cells using ab32522 at 1:100 dilution.

    • Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [E22] (ab32522)
      Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [E22] (ab32522)

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Caspase-7 (red) with ab32522 at a 1/250 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies. 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [E22] (ab32522)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [E22] (ab32522)

      Immunohistochemical analysis of paraffin embedded human skin cancer tissue using ab32522 at 1:50 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-Caspase-7 antibody [E22] (ab32522)
      Immunoprecipitation - Anti-Caspase-7 antibody [E22] (ab32522)

      Purified ab32522 at 1/20 dilution (1µg) immunoprecipitating Caspase-7 in Jurkat whole cell lysate.
      Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg
      Lane 2 (+): ab32522 + Jurkat whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32522 in Jurkat whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 34 kDa

    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 (22)

    Publishing research using ab32522? Please let us know so that we can cite the reference in this datasheet.

    ab32522 has been referenced in 22 publications.

    • Zhao X  et al. Casein Kinase 2-Interacting Protein-1 Alleviates High Glucose-Reduced Autophagy, Oxidative Stress, and Apoptosis in Retinal Pigment Epithelial Cells via Activating the p62/KEAP1/NRF2 Signaling Pathway. J Ophthalmol 2021:6694050 (2021). PubMed: 33628480
    • Gao Z  et al. FK506-binding protein 5 promotes the progression of papillary thyroid carcinoma. J Int Med Res 49:3000605211008325 (2021). PubMed: 33906532
    • Chang Y  et al. MIR205HG facilitates carcinogenesis of lung squamous cell carcinoma in vitro revealed by long noncoding RNA profiling. Acta Biochim Biophys Sin (Shanghai) 52:371-381 (2020). PubMed: 32188965
    • Albahde MAH  et al. CBL0137 administration suppresses human hepatocellular carcinoma cells proliferation and induces apoptosis associated with multiple cell death related proteins. Neoplasma 67:547-556 (2020). PubMed: 32202904
    • Wu SQ  et al. A novel role for Livin in the response to ultraviolet B radiation and pterygium development. Int J Mol Med 45:1103-1111 (2020). PubMed: 32124942
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Immunoprecipitation abreview for Anti-Caspase 7 antibody [E22]

    Average
    Abreviews
    Abreviews
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein A/G
    Sample
    Human Cell lysate - whole cell (MCF7)
    Specification
    MCF7
    Total protein in input
    100 µg
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 05 2013

    Western blot abreview for Anti-Caspase-7 [E22] antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    100 µg
    Gel Running Conditions
    Reduced Denaturing (8)
    Sample
    Human Cell lysate - whole cell (MCF7)
    Specification
    MCF7
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 04 2013

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