Anti-Caspase-3抗体
参阅全部 Caspase-3 一抗
兔多克隆抗体to Caspase-3
Rabbit
Stimulation is required to allow detection of the 17kDa cleaved form of the protein. Please see images below for recommended treatment conditions and positive controls.
ab13847 recognizes a cleaved form of Caspase 3 (~17 kDa) after apoptosis has been induced in wildtype cells and not Caspase 3 knockout cells.
Some customers have used this antibody successfully in IHC-P however our latest tests were unsuccessful and therefore we can no longer guarantee this application. We would recommend ab32351 and ab184787 as an alternative product for this application.
适用于: WBmore details
与反应: Human
预测可用于: Mouse, Rat, Dog, Pig, Xenopus laevis, Drosophila melanogaster, Indian muntjac, Zebrafish, Rhesus monkey, Chinese hamster, Common marmoset, Schmidtea mediterranea, Salvelinus alpinus
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
WB: Hap-1 WT, Human Caspase 3 (active) Recombinant Protein ICC: HeLa cells treated with 1 mM staurosporine
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab13847于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (15) | 1/500. Detects a band of approximately 17, 34 kDa (predicted molecular weight: 17, 34 kDa). |
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SwissProt: P70677 Mouse
SwissProt: P55866 Xenopus laevis
Unigene: 141125 Human
Unigene: 34405 Mouse
Unigene: 10562 Rat
Unigene: 17957 Xenopus laevis
Entrez Gene: 836 Human
Entrez Gene: 12367 Mouse
Entrez Gene: 397720 Xenopus laevis
Entrez Gene: 140621 Zebrafish
Omim: 600636 Human
SwissProt: P42574 Human
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Western blot - Anti-Caspase-3 antibody (ab13847)This image is courtesy of an anonymous Abreview
All lanes : Anti-Caspase-3 antibody (ab13847) at 1/1000 dilution
Lane 1 : Skin (atopic dermatits) whole cell lysate
Lane 2 : Skin (without atopic dermatits) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti rabbit HRP at 1/2000 dilution
Performed under reducing conditions.
Predicted band size: 17, 34 kDa
Observed band size: 30,17 kDawhy is the actual band size different from the predicted?
Additional bands at: 110,130 kDa (possible non-specific binding), 50,70 kDa (possible non-specific binding)
Exposure time: 5 minutes
4-20% (Mini protean gels) non-denaturing conditions
Western blot - Anti-Caspase-3 antibody (ab13847)
Lane 1: Wild-type HAP1 cell lysate + Staurosporine (ab146588) (1μM for 4h)
Lane 2: Wild-type HAP1 cell lysate
Lane 3: Caspase-3 knockout HAP1 cell lysate + Staurosporine (ab146588) (1μM for 4h)
Lane 4: Caspase-3 knockout HAP1 cell lysate
Lanes 1 - 4: Merged signal (red and green). Green - ab13847 observed at 17 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab13847 was shown to recognise Caspase 3 when Caspase 3 knockout samples were used, along with additional cross-reactive bands. Wild-type and Caspase 3 knockout samples (±staurosporine treatment) were subjected to SDS-PAGE. ab13847 and ab8245 (loading control to GAPDH) were diluted to 1/500 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Caspase-3 antibody (ab13847)
All lanes : Anti-Caspase-3 antibody (ab13847) at 1 µg/ml
Lane 1 : Human Caspase 3 (active) Recombinant Protein
Lane 2 : Human Pro Caspase 3 (inactive) Recombinant Protein
Lysates/proteins at 0.1 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 17, 34 kDa
Observed band size: 17,32 kDawhy is the actual band size different from the predicted?
Exposure time: 8 minutes
Caspase 3 exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce large (17kDa) and small (12kDa) subunits. These subunits dimerize to form the active enzyme. ab13847 specifically detects the large active subunit (17kDa) and the inactive pro Caspase 3 (32 kDa).
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 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 ab13847 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
Secondary antibody - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody
