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
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- Frontiers in cellular and infection microbiology 13:12152052023Hydrogel-based 3D human iPSC-derived neuronal culture for the study of rabies virus infection.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPapon Muangsanit,Thanathom Chailangkarn,Nathiphat Tanwattana,Ratjika Wongwanakul,Porntippa Lekcharoensuk,Challika KaewborisuthPubMed 37692167
- Cell death discovery 9:2422023Dysregulation of a lncRNA within the TNFRSF10A locus activates cell death pathways.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTadeusz J Kaczynski,Nadine J Husami,Elizabeth D Au,Michael H FarkasPubMed 37443108
- Bioengineering & translational medicine 8:e104612023Lethal effects of mitochondria via microfluidics.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHyueyun Kim,Young-Ho Ahn,Chang Mo Moon,Jihee Lee Kang,Minna Woo,Minsuk KimPubMed 37206227
- Biological procedures online 24:162022Shrimp miR-965 transfers tumoricidal mitochondria.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHyueyun Kim,Ji Ha Choi,Chang Mo Moon,Jihee Lee Kang,Minna Woo,Minsuk KimPubMed 36289539
- Nature methods 19:1449-14602022Geometric engineering of organoid culture for enhanced organogenesis in a dish.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSunghee Estelle Park,Shawn Kang,Jungwook Paek,Andrei Georgescu,Jeehan Chang,Alex Yoon Yi,Benjamin J Wilkins,Tatiana A Karakasheva,Kathryn E Hamilton,Dan Dongeun HuhPubMed 36280722
- Caspian journal of internal medicine 13:264-2692022Altered serum and cerebrospinal fluid TNF-α, caspase 3, and IL 1β in COVID-19 disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMahtab Ramezani,Omidvar Rezaei,Ilad Alavi Darzam,Mohammadreza Hajiesmaili,Mahdi Amirdosara,Leila Simani,Abbas AliaghaeiPubMed 35872670
- Molecular cell 82:2385-2400.e92022Human NLRP1 is a sensor of pathogenic coronavirus 3CL proteases in lung epithelial cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRémi Planès,Miriam Pinilla,Karin Santoni,Audrey Hessel,Charlotte Passemar,Kenneth Lay,Perrine Paillette,Ana-Luiza Chaves Valadão,Kim Samirah Robinson,Paul Bastard,Nathaniel Lam,Ricardo Fadrique,Ida Rossi,David Pericat,Salimata Bagayoko,Stephen Adonai Leon-Icaza,Yoann Rombouts,Eric Perouzel,Michèle Tiraby,Qian Zhang,Pietro Cicuta,Emmanuelle Jouanguy,Olivier Neyrolles,Clare E Bryant,Andres R Floto,Caroline Goujon,Franklin Zhong Lei,Guillaume Martin-Blondel,Stein Silva,Jean-Laurent Casanova,Céline Cougoule,Bruno Reversade,Julien Marcoux,Emmanuel Ravet,Etienne MeunierPubMed 35594856
- Pharmaceutics 13:2021Antitumor Activity of Nanoparticles Loaded with PHT-427, a Novel AKT/PDK1 Inhibitor, for the Treatment of Head and Neck Squamous Cell Carcinoma.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJoaquín Yanes-Díaz,Raquel Palao-Suay,María Rosa Aguilar,Juan Ignacio Riestra-Ayora,Antonio Ferruelo-Alonso,Luis Rojo Del Olmo,Blanca Vázquez-Lasa,Ricardo Sanz-Fernández,Carolina Sánchez-RodríguezPubMed 34452203
- Journal of Cancer 12:3393-34062021ZFPM2-AS1 transcriptionally mediated by STAT1 regulates thyroid cancer cell growth, migration and invasion via miR-515-5p/TUSC3.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRuizhen Ren,Yuanna Du,Xing Niu,Rukun ZangPubMed 33976749
- Ultrasound in medicine & biology 47:1596-16152021Evaluation of Loading Strategies to Improve Tumor Uptake of Gemcitabine in a Murine Orthotopic Bladder Cancer Model Using Ultrasound and Microbubbles.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJia-Ling Ruan,Richard J Browning,Yesna O Yildiz,Luca Bau,Sukanta Kamila,Michael D Gray,Lisa Folkes,Alix Hampson,Anthony P McHale,John F Callan,Borivoj Vojnovic,Anne E Kiltie,Eleanor StridePubMed 33707089
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture extracts, Tissue Extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 31.25 - 2000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 5.8 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Thiol protease that acts as a major effector caspase involved in the execution phase of apoptosis (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:35338844, PubMed:35446120, PubMed:7596430). Following cleavage and activation by initiator caspases (CASP8, CASP9 and/or CASP10), mediates execution of apoptosis by catalyzing cleavage of many proteins (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:7596430). At the onset of apoptosis, it proteolytically cleaves poly(ADP-ribose) polymerase PARP1 at a '216-Asp-|-Gly-217' bond (PubMed:10497198, PubMed:16374543, PubMed:7596430, PubMed:7774019). Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain (By similarity). Cleaves and activates caspase-6, -7 and -9 (CASP6, CASP7 and CASP9, respectively) (PubMed:7596430). Cleaves and inactivates interleukin-18 (IL18) (PubMed:37993714, PubMed:9334240). Involved in the cleavage of huntingtin (PubMed:8696339). Triggers cell adhesion in sympathetic neurons through RET cleavage (PubMed:21357690). Cleaves and inhibits serine/threonine-protein kinase AKT1 in response to oxidative stress (PubMed:23152800). Acts as an inhibitor of type I interferon production during virus-induced apoptosis by mediating cleavage of antiviral proteins CGAS, IRF3 and MAVS, thereby preventing cytokine overproduction (PubMed:30878284). Also involved in pyroptosis by mediating cleavage and activation of gasdermin-E (GSDME) (PubMed:35338844, PubMed:35446120). Cleaves XRCC4 and phospholipid scramblase proteins XKR4, XKR8 and XKR9, leading to promote phosphatidylserine exposure on apoptotic cell surface (PubMed:23845944, PubMed:33725486). Cleaves BIRC6 following inhibition of BIRC6-caspase binding by DIABLO/SMAC (PubMed:36758104, PubMed:36758106).
Alternative names
CPP32, CASP3, Caspase-3, CASP-3, Apopain, Cysteine protease CPP32, Protein Yama, SREBP cleavage activity 1, CPP-32, SCA-1
What's included?
Recommended products
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
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 31.25 - 2000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Microplate (12 x 8 well strips)
- Sensitivity
- = 5.8 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Cell Extract | n 3 | C.V. 4.2 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Cell Extract | n 5 | C.V. 5.2 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture extracts | Average % = 108 | Range 105 - 115 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Human Cleaved Caspase-3 (Asp175) ELISA Kit (ab220655) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Cleaved Caspase-3 (Asp175) protein in cell culture extracts and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Cleaved Caspase-3 (Asp175) with 2.1 pg/ml sensitivity.
SimpleStep ELISA® technology 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. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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.
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4 product images
ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (ab220655)
Sandwich ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (ab220655)
Comparison of staurosporine treated Jurkat cell extracts collected at different time points during treatment.
Jurkat cells were cultured in the presence of 1 μg/mL staurosporine and collected at time points of 2 hours, 4 hours and 6 hours, or in the presence of staurosporine solvent (mock) collected at 4 hours. The concentration of Active Caspase-3 (Asp175) was measured in three different dilutions of the cell extract samples in duplicate and interpolated from the Active Caspase-3 (Asp175) standard curve. The interpolated dilution factor corrected values are plotted in ng of Active Caspase-3 (Asp175) per mg of extract (mean +/- SD, n=3). The mean Active Caspase-3 (Asp175) concentration was determined to be not detectable in Jurkat (mock treated), 21.16 ng/mg in Jurkat (2 hour), 49.00 ng/mg in Jurkat (4 hour) and 38.79 ng/mg in Jurkat (6 hour) cell extracts.
Sandwich ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (ab220655)
Example of human Active Caspase-3 (Asp175) standard curve in 1X Cell Extraction Buffer PTR.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (ab220655)
Interpolated concentrations of native Active Caspase-3 (Asp175) in staurosporine treated Jurkat cell extract samples collected at different time points.
Jurkat cells were cultured in the presence of 1 μM staurosporine and collected at time points of 2 hours, 4 hours and 6 hours based on a 10 μg/mL extract load. The concentrations of Active Caspase-3 (Asp175) were measured in duplicate and interpolated from the Active Caspase-3 (Asp175) standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Active Caspase-3 (Asp175) concentration was determined to be 206.0 pg/mL in staurosporine treated Jurkat (2 hour), 472.9 pg/mL in staurosporine treated Jurkat (4 hour) and 376.7 pg/mL in staurosporine treated Jurkat (6 hour) cell extracts.
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