AB220655

Human Cleaved Caspase-3 (Asp175) ELISA Kit

Recombinant
SimpleStep
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(16 Publications)

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

View Alternative Names

CPP32, CASP3, Caspase-3, CASP-3, Apopain, Cysteine protease CPP32, Protein Yama, SREBP cleavage activity 1, CPP-32, SCA-1

4 Images

ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (AB220655)

ELISA

Supplier Data

ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (AB220655)

sELISA

Supplier Data

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.

sELISA

Supplier Data

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.

sELISA

Supplier Data

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.

Key facts

Detection method

Colorimetric

Sample types

Cell culture extracts, Tissue Extracts

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 5.8 pg/mL

Range

31.25 - 2000 pg/mL

Assay time

1h 30m

Assay Platform

Microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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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.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Cell Extract", "replicates": 5, "mean": null, "standardDeviation": null, "coefficientOfVariability": "5.2" }, { "reproducibilityType": "Intra", "sample": "Cell Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "4.2" } ]

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Recovery

[ { "sample": "Cell culture extracts", "range": "105 - 115 %", "average": "= 108" } ]

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What's included?

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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

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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.

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Product protocols

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Target data

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 DSG2 in response to apoptosis resulting in a loss of full length DSG2 at desmosome cell junctions and subsequent loss of cell-cell adhesion (PubMed : 17559062). Also cleaves JUP in response to apoptosis (PubMed : 17559062). 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).

See full target information CASP3

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Publications (16)

Recent publications for all applications. Explore the full list and refine your search

International journal of biological sciences 20:6090-6113 PubMed39664583

2024

Repurposing of the Antipsychotic Trifluoperazine Induces SLC7A11/GPX4- Mediated Ferroptosis of Oral Cancer via the ROS/Autophagy Pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Sheng-Chieh Tsai,Po-Chih Chang,Yu Tong Lin,Po-Tsang Huang,Jeff Yi-Fu Chen,Chang-Shen Lin,Bin-Nan Wu,Hui-Min Chang,Wan-Ju Wu,Chi-I Chang,Chien-Hsing Lee

Frontiers in cellular and infection microbiology 13:1215205 PubMed37692167

2023

Hydrogel-based 3D human iPSC-derived neuronal culture for the study of rabies virus infection.

Applications

Unspecified application

Species

Unspecified reactive species

Papon Muangsanit,Thanathom Chailangkarn,Nathiphat Tanwattana,Ratjika Wongwanakul,Porntippa Lekcharoensuk,Challika Kaewborisuth

Cell death discovery 9:242 PubMed37443108

2023

Dysregulation of a lncRNA within the TNFRSF10A locus activates cell death pathways.

Applications

Unspecified application

Species

Unspecified reactive species

Tadeusz J Kaczynski,Nadine J Husami,Elizabeth D Au,Michael H Farkas

Bioengineering & translational medicine 8:e10461 PubMed37206227

2023

Lethal effects of mitochondria via microfluidics.

Applications

Unspecified application

Species

Unspecified reactive species

Hyueyun Kim,Young-Ho Ahn,Chang Mo Moon,Jihee Lee Kang,Minna Woo,Minsuk Kim

Biological procedures online 24:16 PubMed36289539

2022

Shrimp miR-965 transfers tumoricidal mitochondria.

Applications

Unspecified application

Species

Unspecified reactive species

Hyueyun Kim,Ji Ha Choi,Chang Mo Moon,Jihee Lee Kang,Minna Woo,Minsuk Kim

Nature methods 19:1449-1460 PubMed36280722

2022

Geometric engineering of organoid culture for enhanced organogenesis in a dish.

Applications

Unspecified application

Species

Unspecified reactive species

Sunghee Estelle Park,Shawn Kang,Jungwook Paek,Andrei Georgescu,Jeehan Chang,Alex Yoon Yi,Benjamin J Wilkins,Tatiana A Karakasheva,Kathryn E Hamilton,Dan Dongeun Huh

Caspian journal of internal medicine 13:264-269 PubMed35872670

2022

Altered serum and cerebrospinal fluid TNF-α, caspase 3, and IL 1β in COVID-19 disease.

Applications

Unspecified application

Species

Unspecified reactive species

Mahtab Ramezani,Omidvar Rezaei,Ilad Alavi Darzam,Mohammadreza Hajiesmaili,Mahdi Amirdosara,Leila Simani,Abbas Aliaghaei

Molecular cell 82:2385-2400.e9 PubMed35594856

2022

Human NLRP1 is a sensor of pathogenic coronavirus 3CL proteases in lung epithelial cells.

Applications

Unspecified application

Species

Unspecified reactive species

Rémi 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 Meunier

Pharmaceutics 13: PubMed34452203

2021

Antitumor Activity of Nanoparticles Loaded with PHT-427, a Novel AKT/PDK1 Inhibitor, for the Treatment of Head and Neck Squamous Cell Carcinoma.

Applications

Unspecified application

Species

Unspecified reactive species

Joaquín 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-Rodríguez

Journal of Cancer 12:3393-3406 PubMed33976749

2021

ZFPM2-AS1 transcriptionally mediated by STAT1 regulates thyroid cancer cell growth, migration and invasion via miR-515-5p/TUSC3.

Applications

Unspecified application

Species

Unspecified reactive species

Ruizhen Ren,Yuanna Du,Xing Niu,Rukun Zang

View all publications

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Human Cleaved Caspase-3 (Asp175) ELISA Kit

ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (AB220655)

Sandwich ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (AB220655)

Sandwich ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (AB220655)

Sandwich ELISA - Human Cleaved Caspase-3 (Asp175) ELISA Kit (AB220655)

Key facts

Detection method

Sample types

Reacts with

Assay type

Results type

Sensitivity

Range

Assay time

Assay Platform

Complementary Products

Assay Kits

Assay Kits

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Precision

Recovery

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (16)

Product promise