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  1. Link

    caspase-7-antibody-epr22839-12-ab255818.pdf

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

Recombinant Anti-Caspase-7 antibody [EPR22839-12] (ab255818)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (1)

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Western blot - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
  • Western blot - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
  • Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
  • Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
  • Anti-Caspase-7 antibody [EPR22839-12] (ab255818)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22839-12] to Caspase-7
  • Suitable for: Flow Cyt (Intra), WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-Caspase-7 antibody [EPR22839-12]
    See all Caspase-7 primary antibodies
  • Description

    Rabbit monoclonal [EPR22839-12] to Caspase-7
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Untreated wild-type HAP1 whole cell lysate, Wild-type HAP1 treated with 1 M staurosporine for 3 hours whole cell lysate, C6, PC-12, RAW 264.7 whole cell lysate, Untreated HeLa, HeLa treated with 1 M staurosporine for 3 hours whole cell lysate and C2C12 lysate. IHC-P: Human, mouse and rat colon tissues. Flow Cyt (intra): HeLa and C2C12 cells.
  • General notes

    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.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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

    EPR22839-12
  • 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

    • Anti-Caspase-7 antibody [EPR22839-12] - BSA and Azide free (ab256539)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Immunohistochemistry kits

    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human Caspase-7 protein (ab198478)
  • Related Products

    • Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab255818 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
Flow Cyt (Intra)
1/50.
WB
1/1000. Detects a band of approximately 35, 32, 18 kDa (predicted molecular weight: 34 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
Flow Cyt (Intra)
1/50.
WB
1/1000. Detects a band of approximately 35, 32, 18 kDa (predicted molecular weight: 34 kDa).
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC/IF or IP.

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
    • Entrez Gene: 12369 Mouse
    • Entrez Gene: 64026 Rat
    • Omim: 601761 Human
    • SwissProt: P55210 Human
    • SwissProt: P97864 Mouse
    • Unigene: 9216 Human
    • Unigene: 35687 Mouse
    • 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 [EPR22839-12] (ab255818)
      Western blot - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      All lanes : Anti-Caspase-7 antibody [EPR22839-12] (ab255818) at 1/1000 dilution

      Lane 1 : Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : HeLa treated with 1 µM staurosporine for 3 hours whole cell lysate
      Lane 3 : C2C12 (mouse myoblasts myoblast) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 34 kDa
      Observed band size: 18,32,35 kDa why is the actual band size different from the predicted?



      Caspase-7 is activated upon apoptotic induction. Procapase-7 (35 kDa), intermediate caspase-7 (32 kDa) and caspase-7 p20 subunit (18 kDa) are observed. The molecular weight observed is consistent with what has been described in the literature (PMID: 15953353, 18596415).

      Exposure time: Lanes 1-2: 15 seconds; Lane 3: 3 seconds.

      Blocking and Diluting Buffer and concentration: 5% NFDM/TBST.

    • Western blot - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      Western blot - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      All lanes : Anti-Caspase-7 antibody [EPR22839-12] (ab255818) at 1/1000 dilution

      Lane 1 : Untreated wild-type HAP1 whole cell lysate
      Lane 2 : Wild-type HAP1 treated with 1 µM staurosporine for 3 hours whole cell lysate
      Lane 3 : Untreated caspase-7 knockout HAP1 whole cell lysate
      Lane 4 : Caspase-7 knockout HAP1 treated with 1 µM staurosporine for 3 hours whole cell lysate
      Lane 5 : C6 (rat glial tumor glial cell) whole cell lysate
      Lane 6 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
      Lane 7 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 34 kDa
      Observed band size: 32,35 kDa why is the actual band size different from the predicted?


      Exposure time: 26 seconds


      Caspase-7 is activated upon apoptotic induction. Procapase-7 (35 kDa), intermediate caspase-7 (32 kDa) and caspase-7 p20 subunit (18 kDa) are observed. The molecular weight observed is consistent with what has been described in the literature (PMID: 15953353, 18596415). 

      ab255818 was shown to specifically react with Caspase-7 in wild-type HAP1 cells as signal was lost in Caspase-7 knockout cells. Wild-type and Caspase-7 knockout samples were subjected to SDS-PAGE. ab255818 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ instrument using the ECL technique. Blocking and Diluting Buffer and concentration: 5% NFDM/TBST

    • Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized C2C12 (mouse muscle) cells labeling Caspase-7 with ab255818 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      Flow Cytometry (Intracellular) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma) cells labeling Caspase-7 with ab255818 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)

      Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Caspase-7 with ab255818 at 1/2000 dilution (0.28 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat colon (PMID: 30448733) is observed. The section was incubated with ab255818 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)

      Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Caspase-7 with ab255818 at 1/2000 dilution (0.28 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse colon (PMID: 23572575) is observed. The section was incubated with ab255818 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-7 antibody [EPR22839-12] (ab255818)

      Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Caspase-7 with ab255818 at 1/2000 dilution (0.28 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon (PMID: 30448733) is observed. The section was incubated with ab255818 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      Secondary antibody only control/ Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    • Anti-Caspase-7 antibody [EPR22839-12] (ab255818)
      Anti-Caspase-7 antibody [EPR22839-12] (ab255818)

    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

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (1)

    Publishing research using ab255818? Please let us know so that we can cite the reference in this datasheet.

    ab255818 has been referenced in 1 publication.

    • Liu H  et al. Effects of repetitive magnetic stimulation on motor function and GAP43 and 5-HT expression in rats with spinal cord injury. J Int Med Res 48:300060520970765 (2020). PubMed: 33356694

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