Product nameAnti-active Caspase-3 antibody [E83-77]
See all active Caspase-3 primary antibodies
DescriptionRabbit monoclonal [E83-77] to active Caspase-3
SpecificityMouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information This antibody only detects the active (cleaved) form of Caspase-3 and does not recognize the pro form of Caspase-3. PubMedID 19789217 describes the detection of human cells injected into mice. The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway.
Tested applicationsSuitable for: IHC-Fr, WB, ICC/IFmore details
Unsuitable for: Flow Cyt or IP
Species reactivityReacts with: Human
Synthetic peptide within Human active Caspase-3 aa 1-100 (N terminal). The exact sequence is proprietary. A synthetic peptide corresponding to residues following Ser29 of human Caspase 3 (N terminus of p17 subunit).
Database link: P42574
- WB: Wild type HAP1 + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate; Jurkat cell lysate (camptothecin treated); HeLa cell lysate (staurosporine treated). ICC/IF: Hela cells (staurosporine treated); Human Vascular endothelial cells.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
Our Abpromise guarantee covers the use of ab32042 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|WB||1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 32 kDa).
The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway.
|ICC/IF||1/100 - 1/250.|
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
- Apopain antibody
- CASP 3 antibody
- CASP-3 antibody
High-glucose induces apoptosis in human Vascular endothelial cells (VECs).
Apoptotic responses in VEC were analyzed by detection of active-Caspase-3 immunofluorescence using ab32042. Cells were treated with low glucose (LG) or high glucose (HG) for 72 hours before treated with 100 ng/mL bFGF, 1 μM sp600125 (sp), or 1 μM U0126 (U) or 10 μM MnTmPyP for 1 hour. Bar = 100 μm.
Lane 1: Wild type HAP1 + DMSO for 24 hours, whole cell lysate (20 µg)
Lane 2: Wild type HAP1 + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg)
Lane 3: HAP1 CASP3 KO + DMSO for 24 hours, whole cell lysate (20 µg)
Lane 4: HAP1 CASP3 KO + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg)
Lane 5: HeLa + DMSO for 24 hours, whole cell lysate (20 µg)
Lane 6: HeLa + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg)
Lanes 1 - 6: Merged signal (red and green). Green - ab32042 observed at 17 kDa. Red - loading control, ab8245, observed at 130 kDa.
ab32042 was shown to specifically react with CASP3 (Caspase-3) when CASP3 (Caspase-3) knockout samples were used. HAP1 wild-type and CASP3 (Caspase-3) knockout samples were subjected to SDS-PAGE. Ab32042 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Cells were grown to confluency prior to treatment.
Ab32042, at dilution of 1/100, staining HeLa (human epithelial cell line from cervix adenocarcinoma) cells by Immunofluorescence.
Left image: control.
Right image: staurosporine treated.
All lanes : Anti-active Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution
Lane 1 : HeLa Whole Cell Lysate (2 uM Staurosporine, 4Hr) at 20 µg
Lane 2 : HeLa Whole Cell Lysate (untreated) at 20 µg
Lane 3 : Active Caspase 3 (recombinant protein) at 0.1 µg
All lanes : 800CW Goat Anti-Rabbit IgG at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 32 kDa
Additional bands at: 17 kDa (possible mature (processed) protein)
Lane 1 : anti Pro Caspase 3 at 1/10000 dilution
Lane 2 : anti Pro Caspase 3 at 1/10000 dilution
Lanes 3-4 : Anti-active Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution
Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lanes 2 & 4 : Jurkat cell lysate + Camptothecin
Lane 3 : Jurkat cell lysate
Predicted band size: 32 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands.
This product has been referenced in:
- Dai Y et al. Isoquercetin attenuates oxidative stress and neuronal apoptosis after ischemia/reperfusion injury via Nrf2-mediated inhibition of the NOX4/ROS/NF-?B pathway. Chem Biol Interact 284:32-40 (2018). WB . Read more (PubMed: 29454613) »
- Kong D et al. Schizandrin A enhances chemosensitivity of colon carcinoma cells to 5-fluorouracil through up-regulation of miR-195. Biomed Pharmacother 99:176-183 (2018). WB . Read more (PubMed: 29331856) »