Anti-active Caspase-3抗体(ab2302)

概述

  • 产品名称Anti-active Caspase-3抗体
    参阅全部 active Caspase-3 一抗
  • 描述
    兔多克隆抗体to active Caspase-3
  • 特异性The antibody preferentially recognizes the p17 fragment of the active Caspase-3. Under the conditions we tested, the antibody did not detect the precursor form. Under other conditions (such as those used in PMC2206181), this antibody has been found to detect both the pro and active forms of Caspase-3. Although some customers have used this antibody successfully in mouse and rat, we do not batch test in these species and so cannot guarantee that it will work in them.
  • 经测试应用适用于: IHC-FoFr, IHC-P, IHC-Fr, WBmore details
  • 种属反应性
    与反应: Human, Quail
    预测可用于: Mouse, Rat
  • 免疫原

    Synthetic peptide corresponding to Human active Caspase-3. Synthetic peptide mapping to the N-terminus adjacent to the cleavage site of human caspase-3
    Database link: P42574
    (Peptide available as ab38283)

  • 阳性对照
    • Camptothecin (2 µM) treated Jurkat cells. If no signal is observed a time course may be required to identify maximal caspase activity.
  • 常规说明

     

    Caspases are synthesized as inactive pro-enzymes that are processed to active form in cells undergoing apoptosis. Caspase 3 has been extensively studied and implicated to play an important role in apoptosis. Active caspase 3 proteolytically cleaves and activates other caspases, as well as relevant targets in the cells (e.g., PARP). This affinity purified antibody recognizing the active forms of caspase-3 provides a new tool for identifying apoptotic cell populations in both tissue sections and cultured cells.

性能

应用

Our Abpromise guarantee covers the use of ab2302 in the following tested applications.

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

应用 Ab评论 说明
IHC-FoFr Use at an assay dependent concentration. PubMed: 18797916
IHC-P Use a concentration of 5 - 20 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr Use a concentration of 10 - 20 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa.Can be blocked with Human Caspase-3 peptide (ab38283).

靶标

  • 功能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. Triggers cell adhesion in sympathetic neurons through RET cleavage.
  • 组织特异性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.
  • 序列相似性Belongs to the peptidase C14A family.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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 图像

  • IHC image of active caspase-3 staining in human tonsil formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2302, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-active Caspase-3 antibody (ab2302) at 1 µg/ml

    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.

    Additional bands at : 17 kDa (possible mature (processed) protein),24 kDa,35 kDa. We are unsure as to the identity of these extra bands.
  • Immunohistochemistical detection (on formaldehyde/PFA-fixed paraffin-embedded sections) of active Caspase-3 antibody (ab2302) on Quail Tissue sections (Quail E6/7 developing DRGs Saggital section). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins RT. Primary Antibody ab2302 incubated at 1/50 for at RT. Secondary Antibody: Biotin conjugated goat anti rabbit IgG (1/300).

  • ab2302 staining active Caspase-3 in human ulcerative colitis, colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed using the HOPE method and permeabilized. Samples were incubated with primary antibody (1/20) for 1 hour at 25°C. A Biotin-SP-conjugated donkey anti-rabbit IgG polyclonal (1/800) was used as the secondary antibody.

    See Abreview

  • Anti-active Caspase-3 antibody (ab2302) at 1 µg/ml + Camptothecin (2 µM) treated Jurkat cells.
  • Lung sections were stained with caspase antibody ab2302. Caspase-3 positive cells are shown in brown (DAB). This picture was kindly supplied as part of a customer review.

Anti-active Caspase-3 antibody (ab2302)参考文献

This product has been referenced in:
  • Xu TX  et al. Hypoxia responsive miR-210 promotes cell survival and autophagy of endometriotic cells in hypoxia. Eur Rev Med Pharmacol Sci 20:399-406 (2016). WB ; Human . Read more (PubMed: 26914112) »
  • Huang X  et al. Dose-dependent inhibitory effects of zoledronic acid on osteoblast viability and function in vitro. Mol Med Rep 13:613-22 (2016). WB ; Mouse . Read more (PubMed: 26648136) »

See all 78 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Pancreas)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization Yes - Tween 20
Specification Pancreas
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
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提交于 Oct 04 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Brain Tissue and Coronal Suture Cell whole cell ly)
Gel Running Conditions Reduced Denaturing (12% gel)
Loading amount 8 µg
Treatment 10 µM H2O2 for 6 hrs
Specification Brain Tissue and Coronal Suture Cell whole cell ly
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Jun 03 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (PAM212 mouse keratinocyte cell line)
Permeabilization Yes - Yes - 0.2% triton x 100
Specification PAM212 mouse keratinocyte cell line
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative Formaldehyde
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提交于 Jan 06 2016

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (skin)
Permeabilization Yes - 0.2% triton x 100
Specification skin
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10%
Fixative Formaldehyde
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提交于 Dec 02 2015

Application Western blot
Sample Mouse Cell lysate - whole cell (Mouse Glioblastoma (GL261))
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 400000 cells
Treatment DNMT inhibitor
Specification Mouse Glioblastoma (GL261)
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
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提交于 Jun 15 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Pig Tissue lysate - whole (Kidney)
Gel Running Conditions Reduced Denaturing (4-20 % gel)
Loading amount 40 µg
Treatment Ichemia
Specification Kidney
Blocking step BSA as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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提交于 May 27 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citratbuffer
Sample Mouse Tissue sections (Instestine)
Specification Instestine
Permeabilization No
Fixative Methanol
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提交于 Jan 08 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (hepatocytes)
Specification hepatocytes
Treatment 20 ug/ml poly(I:C)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 Jun 30 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step None
Sample Human Tissue sections (Ulcerative colitis, colon)
Specification Ulcerative colitis, colon
Permeabilization Yes - Antibody Diluent (DCS Diagnostics)
Fixative HOPE
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提交于 May 07 2014

Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 25°C
Sample Mouse Tissue sections (uterus)
Specification uterus
Permeabilization No
Fixative Formaldehyde
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Dr. Varkha Agrawal

Verified customer

提交于 Nov 14 2013

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