Key features and details
- Rabbit polyclonal to Cleaved Caspase-3
- Suitable for: IHC-P, WB
- Reacts with: Human, Quail, Recombinant fragment
- Isotype: IgG
Product nameAnti-Cleaved Caspase-3 antibody
See all Cleaved Caspase-3 primary antibodies
DescriptionRabbit polyclonal to Cleaved Caspase-3
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Human, Quail, Recombinant fragment
Predicted to work with: Mouse, Rat
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.
Storage instructionsShipped 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 bufferPreservative: 0.03% Proclin 300
Constituents: PBS, 30% Glycerol (glycerin, glycerine), 0.5% BSA, 0.146% EDTA
Concentration information loading...
PurityImmunogen affinity purified
Primary antibody notesCaspases 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.
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.
|IHC-P||1/10 - 1/20. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.|
|WB||1/50 - 1/500. Detects a band of approximately 17 kDa.|
FunctionInvolved 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.
Tissue specificityHighly 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 similaritiesBelongs to the peptidase C14A family.
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.
- Information by UniProt
- active caspase 3 antibody
- Apopain antibody
- CASP 3 antibody
IHC image of cleaved 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-Cleaved Caspase-3 antibody (ab2302) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (2 uM Staurosporine, 4Hr) at 20 µg
Lane 2 : HeLa whole cell lysate (untreated) at 20 µg
Lane 3 : Cleaved Caspase 3 (recombinant protein) at 0.1 µg
All lanes : 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 cleaved 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 cleaved 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.
Anti-Cleaved Caspase-3 antibody (ab2302) at 1 µg/ml + Camptothecin
(2 µM) treated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
ab2302 has been referenced in 546 publications.
- Li Y et al. LncRNA-LET relieves hypoxia-induced injury in H9c2 cells through regulation of miR-138. J Cell Biochem 121:259-268 (2020). PubMed: 31222827
- Shi CX et al. Sevoflurane attenuates brain damage through inhibiting autophagy and apoptosis in cerebral ischemia-reperfusion rats. Mol Med Rep 21:123-130 (2020). PubMed: 31746402
- Skórkowska A et al. Effect of Combined Prenatal and Adult Benzophenone-3 Dermal Exposure on Factors Regulating Neurodegenerative Processes, Blood Hormone Levels, and Hematological Parameters in Female Rats. Neurotox Res 37:683-701 (2020). PubMed: 31970650
- Ma C et al. The pro-inflammatory effect of NR4A3 in osteoarthritis. J Cell Mol Med 24:930-940 (2020). PubMed: 31701670
- Zhang J et al. Knockdown of RAP2A gene expression suppresses cisplatin resistance in gastric cancer cells. Oncol Lett 19:350-358 (2020). PubMed: 31897147