概述

  • 产品名称Anti-Caspase-2L抗体
    参阅全部 Caspase-2L 一抗
  • 描述
    兔多克隆抗体to Caspase-2L
  • 经测试应用适用于: WB, IHC-Fr, ICC/IF, IHC-P, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Human Caspase-2 aa 300-400 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P42575
    (Peptide available as ab13766)

  • 阳性对照
    • This antibody gave a positive signal in the following cell lysates: Hela (whole cell and nuclear), A431, Jurkat, HEK293 and in the Caspase 2 recombinant protein and in A431 cell line in IF.

性能

应用

Our Abpromise guarantee covers the use of ab2251 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 31 kDa).
IHC-Fr Use at an assay dependent concentration. PubMed: 24223903
ICC/IF Use a concentration of 10 µg/ml.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

靶标

  • 相关性Function: Involved in the activation cascade of caspases responsible for apoptosis execution. Might function by either activating some proteins required for cell death or inactivating proteins necessary for cell survival. Tissue specificity: Expressed at higher levels in the embryonic lung, liver and kidney than in the heart and brain. In adults, higher level expression is seen in the placenta, lung, kidney, and pancreas than in the heart, brain, liver and skeletal muscle. Similarity: Belongs to the peptidase C14A family. Contains 1 CARD domain. PTM: The mature protease can process its own propeptide, but not that of other caspases.
  • 数据库链接
  • 形式Specific for P42575-1; the isoform that has been chosen as the 'canonical' sequence.
  • 别名
    • CASP2 antibody
    • ICH-1L antibody
    • ICH1 antibody
    • NEDD-2 antibody
    • NEDD2 antibody
    • Protease ICH-1 antibody
    see all

Anti-Caspase-2L antibody 图像

  • IHC image of Caspase-2 staining in human hippocampus FFPE section, performed on a Leica BondTM 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 ab2251, 5µg/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.

  • ICC/IF image of ab2251 stained A431 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2251, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, anti-rabbit DyLight® 488 used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-Caspase-2L antibody (ab2251) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : HEK293 Human embryonic kidney cell line Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 31 kDa
    Observed band size : 36 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa,60 kDa,70 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 90 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 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 ab2251 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and  visualised using ECL development solution.

  • Caspase 2 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to Caspase 2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2251.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody anti-Rabbit IgG light chain HRP (ab99697).
    Band: 35kDa: Caspase 2 .

  • Lane 1 : Marker
    Lanes 2 - 7 : Anti-Caspase-2L antibody (ab2251) at 1 µg/ml

    Lane 1 : As above
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab27251) at 20 µg
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg
    Lane 4 : A431 whole cell lysate (ab7909) at 20 µg
    Lane 5 : Jurkat whole cell lysate (ab7899) at 20 µg
    Lane 6 : HEK293 whole cell lysate (ab7902) at 20 µg
    Lane 7 : Recombinant Caspase 2 antibody at 10 µg

    Secondary
    Lanes 2 - 7 : Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution


    Performed under reducing conditions.

    Predicted band size : 31 kDa

    ab2251 detects a band at 35 kDa in cell lysates and a 20 kDa band in the Caspase recombinant protein corresponding to the tagged form.

Anti-Caspase-2L antibody (ab2251)参考文献

This product has been referenced in:

See all 13 Publications for this product

Product Wall

Application Western blot
Loading amount 45 µg
Gel Running Conditions Reduced Denaturing (12% Gel)
Sample Mouse Cell lysate - whole cell (C17.2 cells)
Specification C17.2 cells
Treatment Heat-shock for 2 hours
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
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提交于 Oct 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 45 minute(s) · Concentration: 3% · Temperature: 22°C
Sample Human Tissue sections (Placenta)
Specification Placenta
Permeabilization No
Fixative Paraformaldehyde
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提交于 Sep 30 2013

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