Abcam, ab220655, Human Cleaved Caspase-3 (Asp175) ELISA Kit

Size: 1 x 96Tests
Human Cleaved Caspase-3 (Asp175) ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Cleaved Caspase-3 (Asp175) with a sensitivity of 2.1 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step. - Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader - Design your own immunoassay: we also offer the conjugation-ready antibody pair - Cited in over 10 citations
Key facts
Detection method:Colorimetric,
Sample types:Cell culture extracts, Tissue Extracts,
Reacts with:Human,
Assay type:Sandwich (quantitative),
Sensitivity:= 5.8 pg/mL,
Range:31.25 - 2000 pg/mL,
Assay time:1h 30m,
Assay Platform:Microplate (12 x 8 well strips)

Product details:
Human Cleaved Caspase-3 (Asp175) ELISA Kit ab220655 is a rapid single-wash 90-min Sandwich ELISA to measure Human Cleaved Caspase-3 (Asp175) in cell culture extracts, tissue extracts. This SimpleStep sensitivity is 5.8 pg/mL.
How the assay works
Human Cleaved Caspase-3 (Asp175) SimpleStep ELISA
employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA
plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA
protocol summary in the image section for further details.
Assay Specificity
Our SimpleStep ELISA
kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.
Human Cleaved Caspase-3 (Asp175) ELISA Kit ab220655 protocol summary
1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm
Caspase-3 is a cytoplasmic cysteine protease involved in the activation cascade of caspases responsible for execution of apoptosis. At the onset of apoptosis caspase-3 cleaves and activates caspase-6, -7 and -9. It also cleaves poly (ADP-ribose) polymerase (PARP). Caspase-3 cleaves and activates sterol regulatory element binding proteins (SREBPs). Caspase-3 is involved in the cleavage of huntingtin. Caspase-3 is expressed in an inactive pro-form (pro caspase-3). In apoptosis, the pro caspase-3 is activated by proteolytic cleavages at Asp28-Ser29 and Asp175-Ser176 bonds catalyzed by granzyme B, caspase-6, caspase-8, caspase-9 and caspase-10, generating two subunits p17 and p12 that are assembled into heterotetrameric active enzyme. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. The pro-form and the active form are useful biomarkers of apoptosis.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
The Cleaved Caspase-3 also known as active caspase 3 plays an important role in the execution phase of cell apoptosis. The caspase 3 molecule has a molecular weight of approximately 32 kDa. It undergoes cleavage into subunits that are important for its activation marking its conversion into the cleaved form. Cleaved caspase is widely expressed in various tissues with a notable presence in the cytoplasm of cells undergoing apoptotic processes. Cleavage of caspase 3 results in the formation of a functional enzyme that participates in the degradation of cellular components during apoptosis.
Biological function summary
Caspase 3 acts as an executioner enzyme in the process of programmed cell death. Its cleavage from the inactive zymogen form into the active form triggers apoptotic pathways ensuring the removal of damaged or unnecessary cells. Cleaved caspase operates within a proteolytic cascade often in conjunction with other caspases such as caspase 9. Once activated caspase 3 targets and cleaves specific cellular substrates contributing to the characteristic morphological changes and DNA fragmentation associated with apoptosis.
Pathways
Cleaved caspase 3 is significant within the intrinsic and extrinsic apoptotic pathways. It acts downstream of initiator caspases such as caspase 9 in the intrinsic pathway where its activation involves mitochondrial signals. In the extrinsic pathway it closely associates with caspase 8 mediating cell death signals from extracellular triggers. These pathways ensure that cleaved caspase 3 fulfills its role as a critical mediator of apoptosis linking upstream death signals to cellular dismantling during the apoptosis process.
Cleaved caspase 3 connects to conditions such as cancer and neurodegenerative diseases. In cancer dysregulation of caspase 3 activity may lead to evasion of apoptosis facilitating tumor progression. Therapeutic approaches aim to restore its apoptotic functions in cancer cells. In neurodegenerative diseases like Alzheimer's disease abnormal activation of cleaved caspase 3 contributes to neuronal loss. It interacts with proteins like amyloid precursor protein promoting the pathological changes associated with this disorder. Understanding these interactions provides insights into potential therapeutic avenues.