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详细说明
概述
产品名称Anti-Caspase-9抗体[E23]
描述
兔单克隆抗体[E23] to Caspase-9
特异性This antibody should recognise both the pro-[40kDa] form and p35 cleaved form of Caspase-9.
经测试应用WB,IHC-P,Flow Cyt,IP,ICC/IF
种属反应性
与反应: Human
免疫原
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Caspase-9 aa 250-350.
Database link: P55211阳性对照
WB: Jurkat and HeLa cell lysates. IHC-P: Human skeletal muscle and cervical carcinoma tissues. ICC/IF: HepG2 cells. Flow Cyt: K562 cells. IP: HeLa whole cell lysate.
常规说明
This product is a recombinant rabbit monoclonal antibody.
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.
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.
A trial size is available to purchase for this antibody.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information
性能
形式Liquid
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
存储溶液pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA纯度Protein A purified
克隆单克隆
克隆编号E23
同种型IgG
研究领域
Cancer
Cell Death
Apoptosis
Metabolism
Cancer
Cell Death
Apoptosis
Apoptotic Markers
Cytochrome C
Cancer
Cell Death
Apoptosis
Apoptotic Markers
Caspases
Metabolism
Pathways and Processes
Metabolism processes
Apoptosis
Cancer
Invasion/microenvironment
Apoptosis
Cytochrome C
Cancer
Invasion/microenvironment
Apoptosis
Caspases
Cell Biology
Apoptosis
Intracellular
Caspases etc
Caspases
Anti-Caspase-9 antibody [E23] 图像
![Western blot - Anti-Caspase-9 antibody [E23] (ab32539)](http://img.lianshimall.com/statics/attachment/goods/pl20160426/abcamMainImgPrimary/detail/ab3/ab32539WB1.jpg)
Western blot - Anti-Caspase-9 antibody [E23] (ab32539)
All lanes : Anti-Caspase-9 antibody [E23] (ab32539) at 1/1000 dilution (purified)
Lane 1 : HeLa whole cell lysate - treated with Camptothecin
Lane 2 : HeLa whole cell lysate - untreated
Lysates/proteins at 10 µg per lane.
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size : 46 kDa
Observed band size : 35,46 kDa (why is the actual band size different from the predicted?)
Blocking and dilution buffer: 5% NFDM/TBST.
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] (ab32539)](http://img.lianshimall.com/statics/attachment/goods/pl20160426/abcamMainImgPrimary/detail/ab3/ab32539HC1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] (ab32539)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeltal muscle tissue labelling Caspase-9 with purified ab32539 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
![Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [E23] (ab32539)](http://img.lianshimall.com/statics/attachment/goods/pl20160426/abcamMainImgPrimary/detail/ab3/ab32539IF1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [E23] (ab32539)
Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Caspase-9 with purified ab32539 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
![Flow Cytometry - Anti-Caspase-9 antibody [E23] (ab32539)](http://img.lianshimall.com/statics/attachment/goods/pl20160426/abcamMainImgPrimary/detail/ab3/ab32539FC1.jpg)
Flow Cytometry - Anti-Caspase-9 antibody [E23] (ab32539)
Flow Cytometry analysis of K562 cells labelling Caspase-9 with purified ab32539 at 1/250 (red). Cells were fixed with 100% methanol. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
![Immunoprecipitation - Anti-Caspase-9 antibody [E23] (ab32539)](http://img.lianshimall.com/statics/attachment/goods/pl20160426/abcamMainImgPrimary/detail/ab3/ab32539IP1.jpg)
Immunoprecipitation - Anti-Caspase-9 antibody [E23] (ab32539)
ab32539 (purified) at 1/80 immunoprecipitating Caspase-9 in HeLa whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10µg)
Lane 2 (+): ab32539 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32539 in HeLa whole cell lysate.
For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
![Western blot - Anti-Caspase-9 antibody [E23] (ab32539)](http://img.lianshimall.com/statics/attachment/goods/pl20160426/abcamMainImgPrimary/detail/ab3/ab32539539.jpg)
Western blot - Anti-Caspase-9 antibody [E23] (ab32539)
All lanes : Anti-Caspase-9 antibody [E23] (ab32539) at 1/20000 dilution (unpurified)
Lane 1 : Jurkat cell lysate - untreated
Lane 2 : Jurkat cell lysate - treated with Camptothecin
Predicted band size : 46 kDa
Observed band size : 48 kDa (why is the actual band size different from the predicted?)
Additional bands at : 28 kDa. We are unsure as to the identity of these extra bands.Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] (ab32539)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling Caspase 9 with unpurified ab32539 at a dilution of 1/50.
![Flow Cytometry - Anti-Caspase-9 antibody [E23] (ab32539)](http://img.lianshimall.com/statics/attachment/goods/pl20160426/abcamMainImgPrimary/detail/ab3/ab32539392.jpg)
Flow Cytometry - Anti-Caspase-9 antibody [E23] (ab32539)
Overlay histogram showing K562 cells stained with unpurified ab32539 (red line). The cells were fixed with 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 (unpurified ab32539, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was anti-rabbit DyLight® 488 (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in K562 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.
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