Anti-Caspase-3 antibody (ab13847)
Key features and details
- Rabbit polyclonal to Caspase-3
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
-
Product name
Anti-Caspase-3 antibody
See all Caspase-3 primary antibodies -
Description
Rabbit polyclonal to Caspase-3 -
Host species
Rabbit -
Specificity
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.
-
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Dog, Pig, Xenopus laevis, Drosophila melanogaster, Indian muntjac, Zebrafish, Rhesus monkey, Chinese hamster, Common marmoset, Schmidtea mediterranea, Salvelinus alpinus
-
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Hap-1 WT, Human Caspase 3 (active) Recombinant Protein ICC: HeLa cells treated with 1 mM staurosporine
-
General notes
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
Properties
-
Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
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. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
-
Assay kits
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab13847 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | (9) |
Use a concentration of 5 µg/ml.
|
| WB | (15) |
1/500. Detects a band of approximately 17, 34 kDa (predicted molecular weight: 17, 34 kDa).
|
| Notes |
|---|
|
ICC/IF
Use a concentration of 5 µg/ml. |
|
WB
1/500. Detects a band of approximately 17, 34 kDa (predicted molecular weight: 17, 34 kDa). |
Target
-
Function
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. -
Tissue specificity
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. -
Sequence similarities
Belongs to the peptidase C14A family. -
Post-translational
modificationsCleavage 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. -
Cellular localization
Cytoplasm. - Information by UniProt
-
Database links
- Entrez Gene: 836 Human
- Entrez Gene: 12367 Mouse
- Entrez Gene: 397244 Pig
- Entrez Gene: 25402 Rat
- Entrez Gene: 397720 Xenopus laevis
- Entrez Gene: 140621 Zebrafish
- Omim: 600636 Human
- SwissProt: P42574 Human
see all -
Alternative names
- A830040C14Rik antibody
- Apopain antibody
- CASP 3 antibody
see all
Images
-
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 lysateLanes 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 kDa why is the actual band size different from the predicted?
Exposure time: 8 minutesCaspase 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
-
Immunocytochemistry/ Immunofluorescence - Anti-Caspase-3 antibody (ab13847)This image is courtesy of Roberto Giambruno, Marilena Ciciarello and Patrizia LaviaHeLa cells were fixed for 10 minutes at room temperature in 3.7% PFA and permeabilised in 0.1% Triton X-100/PBS then incubated with ab13847 (5µg/ml) for 1 hour at room temperature. The top panel shows control cells treated with DMSO. The bottom panel shows HeLa cells treated with 1 mM staurosporine (ab146588) for 4 hours to induce caspase-3 activation. ab13847 staining is shown in green and counterstaining with DAPI is shown in blue. 100x magnification.
The image shows the staining with ab13847 is very faint in the untreated control cultures, but very bright after activation of capsase-3 by treatment with the staurosporine . (N.B. in these cultures the nuclei are apoptotic).
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (793)
ab13847 has been referenced in 793 publications.
- Liu Y et al. Paeonol inhibits apoptosis of vascular smooth muscle cells via up-regulation of autophagy by activating class III PI3K/Beclin-1 signaling pathway. Life Sci 264:118714 (2021). PubMed: 33157088
- Jin P et al. Salidroside inhibits apoptosis and autophagy of cardiomyocyte by regulation of circular RNA hsa_circ_0000064 in cardiac ischemia-reperfusion injury. Gene 767:145075 (2021). PubMed: 32858179
- Xie Y et al. miR-302e Suppresses Glioma Progression by Targeting VEGFA. Cancer Manag Res 12:10965-10974 (2020). PubMed: 33154675
- Zhou H et al. Ginseng polysaccharide inhibits MDA-MB-231 cell proliferation by activating the inflammatory response. Exp Ther Med 20:229 (2020). PubMed: 33149784
- Wang B et al. Butorphanol Inhibits the Malignant Biological Behaviors of Ovarian Cancer Cells via Down-Regulating the Expression of TMEFF1. Onco Targets Ther 13:10973-10981 (2020). PubMed: 33149612