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 treated with 1 µM staurosporine for 4 hours whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 32 kDa
Observed MW: 17, 19 kDa
Cleaved Caspase-3 (Asp175) Recombinant Rabbit mAb (S-1406-409)
Cleaved Caspase-3 (Asp175) Recombinant Rabbit mAb (S-1406-409)
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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 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm |
| Accession | P42574 |
| Clone Number | S-1406-409 |
| 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 | 1:250 | Hu, Rt |
| 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.
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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 NIH/3T3 whole cell lysate 20 µg
Lane 2: NIH/3T3 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: 32 kDa
Observed MW: 19 kDa
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 C6 whole cell lysate 20 µg
Lane 2: C6 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: 32 kDa
Observed MW: 17 kDa
This blot was developed with high sensitivity substrate
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human kidney. Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/250 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 tonsil. Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/250 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 cervical squamous cell carcinoma. Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/250 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 rat cerebral cortex. Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/250 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 rat colon. Anti-Cleaved Caspase-3 (Asp175) antibody was used at 1/250 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 whole cell lysate treated with 1uM staurosporine for 4 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).
