製品の概要

  • 製品名Anti-Caspase-3 antibody [E87]
    Caspase-3 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [E87] to Caspase-3
  • 特異性This antibody is specific for the pro form and the p17 cleaved form of human Caspase-3.
  • アプリケーション適用あり: ICC/IF, WB, IHC-P, Flow Cyt, IPmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Caspase-3 aa 50-150.
    Database link: P42574

  • ポジティブ・コントロール
    • WB: Jurkat cell lysate. IHC-P: Cervical carcinoma tissue slides. Flow Cyt: HeLa cells.
  • 特記事項

    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.

    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.

    A trial size is available to purchase for this antibody.

    Rat reactivity: We have preliminary internal testing data indicating this antibody does not react with mouse caspase 3. Internal testing and customer reports also indicate non-reactivity with rat capsase 3, contrary to earlier positive reports. Please contact us for more information.

    Alternative versions available:

    Anti-Caspase-3 antibody (Biotin) [E87] (ab195905)

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32351 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF 1/500.

For unpurified, use 1/25.

WB 1/5000. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa).
IHC-P 1/25 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/180 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/10 - 1/50.

ターゲット情報

  • 機能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.
  • 組織特異性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.
  • 配列類似性Belongs to the peptidase C14A family.
  • 翻訳後修飾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.
  • 細胞内局在Cytoplasm.
  • Information by UniProt
  • 参照データベース
  • 別名
    • A830040C14Rik antibody
    • Apopain 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
    • CC3 antibody
    • CPP-32 antibody
    • CPP32 antibody
    • CPP32B antibody
    • Cysteine protease CPP32 antibody
    • EC 3.4.22.56 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
    • SREBP cleavage activity 1 antibody
    • Yama antibody
    see all

Anti-Caspase-3 antibody [E87] 画像

  • All lanes : Anti-Caspase-3 antibody [E87] (ab32351) at 1/5000 dilution (purified)

    Lane 1 : untreated Jurkat cell lysate
    Lane 2 : Jurkat treated with staurosporine

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 32 kDa
    Observed band size : 17,35 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-Caspase-3 antibody [E87] (ab32351) at 1/5000 dilution (purified)

    Lane 1 : Ramos cell lysate
    Lane 2 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescence staining of Jurkat cells with purified ab32351 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32351 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical staining of paraffin embedded human tonsil with purified ab32351 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • ab32351 (purified) at 1/50 immunoprecipitating Cullin 1 in 10 μg HeLa whole cell lysate (Lanes 1 and 2, observed at 35 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP (HRP) (ab131366) was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
  • Overlay histogram showing Ramos cells fixed in 4% PFA and stained with purified ab32351 at a dilution of 1 in 180 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).


  • Predicted band size : 32 kDa

    Carried out with unpurified antibody. Lane 1 = Caspase 3 protein (Active) (ab52314) ? 20 ng. Lane 2 = Caspase 9 protein (Active) (ab52203) ? 20 ng. Lane 3 = Extract of HeLa cells treated with vehicle (ab136806) ? 20 ug. Lane 4 = Extract of HeLa cells treated with staurosporine (ab136806) ? 20 ug. SDS PAGE performed under reducing conditions (100 mM DTT ? Sample heated at 50?C). Primary : Lanes 1-4: Anti Caspase 3 antibody (ab32351) at 1:1000 dilution. Secondary : Lanes 1-4: Goat anti rabbit IgG(H&L)-HRP at 1:10000. Development: ECL for 10 min exposure. Blocking: in 5% Milk + PBS overnight at 4 C. Primary antibody: in 5% Milk + PBS for 2 hours at RT. Secondary antibody: in 5% Milk + PBS for 2 hours at RT. Predicted band size : 32 kDa and 17 kDa. Observed band size : 32 kDa and 17 kDa.

  • Unpurified ab32351, at a 1/25 dilution, staining Capase-3 in paraffin embedded human cervical carcinoma tissue by Immunohistochemistry.

  • Overlay histogram showing HeLa cells stained with unpurfied ab32351 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32351, 1/1000 dilution) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Anti-Caspase-3 antibody [E87] (ab32351) 使用論文

This product has been referenced in:
  • He X  et al. Functional repair of p53 mutation in colorectal cancer cells using trans-splicing. Oncotarget 6:2034-45 (2015). Human . Read more (PubMed: 25576916) »
  • Xu Y  et al. Proteasome inhibitor lactacystin enhances cisplatin cytotoxicity by increasing endoplasmic reticulum stress-associated apoptosis in HeLa cells. Mol Med Rep 11:189-95 (2015). Read more (PubMed: 25323748) »

See all 24 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (RAMOS cells)
Specification RAMOS cells
Treatment with or without TGFbeta (50pM, 24h)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 May 07 2014

Application Western blot
Sample Human Cell lysate - whole cell (N87 cell line)
Gel Running Conditions Reduced Denaturing (4-20% Tris Glycin gel)
Loading amount 800000 cells
Specification N87 cell line
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Aug 03 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 0.05% trypsin
Sample Mouse Tissue sections (prostate)
Specification prostate
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Dec 05 2014

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Goat serum 5% +BSA1% as blocking agent for 2 hour(s) and 0 minute(s) · Temperature: 23°C
Antigen retrieval step None
Sample Rat Tissue sections (brain)
Specification brain
Permeabilization Yes - triton 0.2%
Fixative Paraformaldehyde
Username

Ms. Francoise Geffroy

Verified customer

投稿 Sep 16 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample African Green Monkey Cell lysate - whole cell (COS-1 cells)
Specification COS-1 cells
Treatment with or without TGFbeta (50pM, 24h)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 May 07 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (NMuMG cells)
Specification NMuMG cells
Treatment with or without TGFbeta (50pM, 24h)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 May 07 2014

This antibody will detect both the pro-form and p17 cleaved-form of Caspase-3 and has been shown to work well in Western blotting with Jurkat cell lysate.

The product I would like to suggest is the following:

Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab97051)

http://www.abcam.com/index.html?datasheet=97051 (or use the following: http://www.abcam.com/index.html?data...

Read More

Thank you for your inquiry.
I am happy to confirm that all these antibodies are tested and guaranteed to work with human samples.
You are correct that we do not have any reports yet about the use of these antibodies in muscle. I assume that i...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization No
Username

Dr. Kirk McManus

Verified customer

投稿 May 31 2012

1-10 of 16 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"