WB result of Cleaved Caspase-3 (Asp175) Recombinant Rabbit mAb
Primary antibody: Cleaved Caspase-3 (Asp175) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa starve overnight, then treated with 1 μM Staurosporine for 3 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 31 kDa
Observed MW: 17, 19 kDa
Cleaved Caspase-3 (Asp175) Recombinant Rabbit mAb (S-2817)
Cleaved Caspase-3 (Asp175) Recombinant Rabbit mAb (S-2817)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Cleaved Caspase-3 (Asp175) |
| Synonyms | Caspase-3; CASP-3; Apopain; Cysteine protease CPP32 (CPP-32); Protein Yama; SREBP cleavage activity 1 (SCA-1); CPP32; CASP3 |
| Location | Cytoplasm |
| Accession | P42574 |
| Clone Number | S-2817 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms, Rt |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms, Rt |
| IHC-P | 1:500 | Hu, Ms |
| ICC | 1:500 | Hu |
Background
Cleaved Caspase-3 (Asp175) is the ~17/19 kDa proteolytically activated form of human caspase-3 that, once cleaved after Asp175-Ser176 during apoptosis, loses its N-terminal pro-domain, assembles into active hetero-tetramers (p17/p12) and relentlessly hydrolyzes Asp-x bonds in >100 cytoplasmic, nuclear and cytoskeletal substrates—such as PARP, ICAD, ROCK1 and gelsolin—thereby dismantling cellular architecture, inactivating DNA repair, triggering chromatin condensation, membrane blebbing and phosphatidyl-serine exposure, and thus serving as the pivotal executioner caspase whose appearance, detected by the widely used “Asp175” neo-epitope antibody, is universally recognized as an irreversible biochemical signature of cells committed to programmed cell death.
Picture
Picture
Western Blot
WB result of Cleaved Caspase-3 (Asp175) Recombinant Rabbit mAb
Primary antibody: Cleaved Caspase-3 (Asp175) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated RAW264.7 whole cell lysate 20 µg
Lane 2: RAW264.7 treated with 1 μM Staurosporine for 3 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 31 kDa
Observed MW: 17, 19 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human tonsil. Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human endometrial cancer. Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse embryo. Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/500 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC analysis of HeLa cells starve overnight then treated with Staurosporine (1μM,3 hours) (top panel) and untreated HeLa cells (below panel). Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
