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Human Active Caspase-3 Quantikine ELISA Kit

Catalog #: KM300
27 Citations Datasheet
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Human Active Caspase-3 ELISA Standard Curve
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Citations (27)
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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human Active Caspase-3 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Extracts (100 uL)
Sensitivity
0.1 ng/mL
Assay Range
0.3 - 20 ng/mL (Cell Extracts)
Specificity
Biotin-ZVKD-fmk modified natural and biotin-D-fmk modified recombinant active Caspase-3. This assay also detects mouse active Caspase-3.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human active Caspase-3 Immunoassay is a 3.5 hour solid phase ELISA designed to measure human active caspase-3 in cell extracts. It contains biotinylated-inhibitor derivatized recombinant caspase-3 and a monoclonal antibody raised against caspase-3. Results obtained using natural active caspase-3 covalently bound to biotin-ZVKD-fmk inhibitor in apoptotic cell extracts showed linear curves that were parallel to the Quantikine kit standard. These results indicate that this kit can be used to determine relative mass values for natural active caspase-3.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Extracts

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 1.06 3.09 7.7 1.06 3.27 8.25
Standard Deviation 0.05 0.16 0.2 0.07 0.19 0.5
CV% 4.7 5.2 2.6 6.6 5.8 6.1

Recovery

The recovery of active caspase-3 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Calibrator Diluent RD5-20 (1X) (n=4) 99 98-100
Extraction Buffer (1X) (n=4) 98 97-101
Extraction Buffer (1X) (n=4) 110 104-114
Extracts from Jurkat Cells Treated w/Staurosporine (n=4) 92 87-95

Linearity

To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of active caspase-3 in each matrix were diluted with Calibrator Diluent and then assayed. Results from typical sample dilutions are
Human Active Caspase-3 ELISA Linearity

Scientific Data

N/A Human Active Caspase-3 ELISA Standard Curve View Larger

Human Active Caspase-3 ELISA Standard Curve

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Product Datasheet

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Caspase-3

Caspases are a family of cytosolic aspartate-specific cysteine proteases involved in the initiation and execution of apoptosis. They are expressed as latent zymogens and are activated by an autoproteolytic mechanism or by processing by other proteases (frequently other caspases). Human caspases can be subdivided into three functional groups: cytokine activation (caspase-1, -4, -5, and -13), apoptosis initiation (caspase-2, -8, -9, -and -10), and apoptosis execution (caspase-3, -6, and -7).

Caspases are regulated by a variety of stimili, including APAF1, CFLAR/FLIP, NOL3/ARC, and members of the inhibitor of apoptosis (IAP) family such as BIRC1/NAIP, BIRC2/cIAP-1, BIRC3/cIAP-2, BIRC4/XIAP, BIRC5/Survivin, and BIRC7/Livin. IAP activity is modulated by DIABLO/SMAC or PRSS25/HTRA2/Omi. Cell-permeable and irreversible peptide inhibitors are also available for different caspases.

Entrez Gene IDs:
836 (Human); 12367 (Mouse)
Alternate Names:
Apopain; apoptosis-related cysteine protease; CASP3; CASP-3; caspase 3, apoptosis-related cysteine peptidase; Caspase3; Caspase-3; CPP32; CPP-32; CPP32B; CPP32SREBP cleavage activity 1; Cysteine protease CPP32; EC 3.4.22; EC 3.4.22.56; LICE-1; PARP cleavage protease; procaspase3; Protein Yama; SCA-1; YAMA
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 100 µL Standard, Control, or Sample
  3.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  4.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    5. 100 µL Conjugate
  5.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  6.   Aspirate and wash 5 times.
    7. 100 µL Substrate Solution
  7.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    8. 100 µL Stop Solution
  8.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Active Caspase-3 Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

27 Citations: Showing 1 - 10
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  1. Regulatory T Cell Apoptosis during Preeclampsia May Be Prevented by Gal-2
    Authors: S Meister, L Hahn, S Beyer, M Mannewitz, C Perleberg, K Schnell, D Anz, S Corradini, E Schmoeckel, D Mayr, U Hasbargen, A Zati Zehni, S Mahner, U Jeschke, T Kolben
    International Journal of Molecular Sciences, 2022-02-07;23(3):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Obesity alters ovarian folliculogenesis through disrupted angiogenesis from increased IL-10 production
    Authors: PK Yang, CH Chou, CC Huang, WF Wen, HF Chen, CT Shun, HN Ho, MJ Chen
    Molecular Metabolism, 2021-02-13;49(0):101189.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. New cyanopyridine-based scaffold as PIM-1 inhibitors and apoptotic inducers: Synthesis and SARs study
    Authors: AM Farrag, MH Ibrahim, ABM Mehany, MMF Ismail
    Bioorg Chem, 2020-10-13;105(0):104378.
    Species: Human
    Sample Types: Cell Lysates
  4. Design and synthesis of novel tranilast analogs: Docking, antiproliferative evaluation and in-silico screening of TGF&betaR1 inhibitors
    Authors: MMF Ismail, HSA El-Zahabi, RS Ibrahim, ABM Mehany
    Bioorg Chem, 2020-10-12;105(0):104368.
    Species: Human
    Sample Types: Cell Lysates
  5. Role of Bee Venom and Melittin on Restraining Angiogenesis and Metastasis in gamma-Irradiated Solid Ehrlich Carcinoma-Bearing Mice
    Authors: NM El Bakary, AZ Alsharkawy, ZA Shouaib, EMS Barakat
    Integr Cancer Ther, 2020-01-01;19(0):1534735420944.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. Apoptosis: A target for anticancer therapy with novel cyanopyridines
    Authors: MMF Ismail, AM Farrag, MF Harras, MH Ibrahim, ABM Mehany
    Bioorg. Chem., 2019-12-05;94(0):103481.
    Species: Human
    Sample Types: Cell Lysates
  7. Immunomodulatory effect of mancozeb, chlorothalonil, and thiophanate methyl pesticides on macrophage cells
    Authors: GCC Weis, CE Assmann, FC Cadoná, BDSR Bonadiman, AO Alves, AK Machado, MMMF Duarte, IBM da Cruz, IH Costabeber
    Ecotoxicol. Environ. Saf., 2019-07-09;182(0):109420.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. Hypomorphic mutations of TRIP11 cause odontochondrodysplasia
    Authors: A Wehrle, TM Witkos, S Unger, J Schneider, JA Follit, J Hermann, T Welting, V Fano, M Hietala, N Vatanavich, K Schoner, J Spranger, M Schmidts, B Zabel, GJ Pazour, A Bloch-Zupa, G Nishimura, A Superti-Fu, M Lowe, E Lausch
    JCI Insight, 2019-02-07;4(3):.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Protective effect of hydroxysafflor yellow A against acute kidney injury via the TLR4/NF-?B signaling pathway
    Authors: J Bai, J Zhao, D Cui, F Wang, Y Song, L Cheng, K Gao, J Wang, L Li, S Li, Y Jia, A Wen
    Sci Rep, 2018-06-15;8(1):9173.
    Species: Rat
    Sample Types: Serum
  10. CCL2 influences the sensitivity of lung cancer A549 cells to docetaxel
    Authors: T Wang, Q Zhan, X Peng, Z Qiu, T Zhao
    Oncol Lett, 2018-05-22;16(1):1267-1274.
    Species: Human
    Sample Types: Cell Lysates
  11. Ghrelin therapy mitigates bone marrow injury and splenocytopenia by sustaining circulating G-CSF and KC increases after irradiation combined with wound
    Authors: JG Kiang, MN Anderson, JT Smith
    Cell Biosci, 2018-04-05;8(0):27.
    Species: Mouse
    Sample Types: Serum
  12. Beneficial Effect of Jojoba Seed Extracts on Hyperglycemia-Induced Oxidative Stress in RINm5f Beta Cells
    Authors: S Belhadj, O Hentati, G Hamdaoui, K Fakhreddin, E Maillard, S Dal, S Sigrist
    Nutrients, 2018-03-20;10(3):.
    Species: Rat
    Sample Types: Cell Lysates
  13. A Novel Role of SIRT1/ FGF-21 in Taurine Protection Against Cafeteria Diet-Induced Steatohepatitis in Rats
    Authors: AH Abd Elwaha, BK Ramadan, MF Schaalan, AM Tolba
    Cell. Physiol. Biochem., 2017-09-22;43(2):644-659.
    Species: Rat
    Sample Types: Tissue Homogenates
  14. Methyltransferase G9a promotes cervical cancer angiogenesis and decreases patient survival
    Authors: RJ Chen, CT Shun, ML Yen, CH Chou, MC Lin
    Oncotarget, 2017-07-07;8(37):62081-62098.
    Species: Human
    Sample Types: Cell Lysates
  15. Bone marrow-derived mesenchymal stem cells mitigate caspase-3 and 8-hydroxy proline induced via ?-adrenergic agonist in pulmonary injured rats
    Authors: MO Kadry, RM Abdel-Mege
    J. Biochem. Mol. Toxicol, 2017-03-07;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
  16. Didox and resveratrol sensitize colorectal cancer cells to doxorubicin via activating apoptosis and ameliorating P-glycoprotein activity
    Sci Rep, 2016-11-14;6(0):36855.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Indole-3- carbinol enhances sorafenib cytotoxicity in hepatocellular carcinoma cells: A mechanistic study
    Sci Rep, 2016-09-09;6(0):32733.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Decidualization and syndecan-1 knock down sensitize endometrial stromal cells to apoptosis induced by embryonic stimuli.
    Authors: Boeddeker S, Baston-Buest D, Fehm T, Kruessel J, Hess A
    PLoS ONE, 2015-04-01;10(4):e0121103.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. A new biocompatible and antibacterial phosphate free glass-ceramic for medical applications.
    Authors: Cabal B, Alou L, Cafini F, Couceiro R, Sevillano D, Esteban-Tejeda L, Guitian F, Torrecillas R, Moya J
    Sci Rep, 2014-06-25;4(0):5440.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Inhibition of nuclear factor-kappa B activation decreases survival of Mycobacterium tuberculosis in human macrophages.
    Authors: Bai X, Feldman N, Chmura K, Ovrutsky A, Su W, Griffin L, Pyeon D, McGibney M, Strand M, Numata M, Murakami S, Gaido L, Honda J, Kinney W, Oberley-Deegan R, Voelker D, Ordway D, Chan E
    PLoS ONE, 2013-04-25;8(4):e61925.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. miR-125b develops chemoresistance in Ewing sarcoma/primitive neuroectodermal tumor.
    Authors: Iida, Keiichir, Fukushi, Jun-Ichi, Matsumoto, Yoshihir, Oda, Yoshinao, Takahashi, Yusuke, Fujiwara, Toshifum, Fujiwara-Okada, Yuko, Hatano, Mihoko, Nabashima, Akira, Kamura, Satoshi, Iwamoto, Yukihide
    Cancer Cell Int, 2013-03-04;13(1):21.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Cyclopamine and jervine induce COX-2 overexpression in human erythroleukemia cells but only cyclopamine has a pro-apoptotic effect.
    Authors: Ghezali L, Leger D, Limami Y, Cook-Moreau J, Beneytout J, Liagre B
    Exp Cell Res, 2013-01-25;319(7):1043-53.
    Species: Human
    Sample Types: Cell Lysates
  23. A systems approach identifies HIPK2 as a key regulator of kidney fibrosis.
    Authors: Jin Y, Ratnam K, Chuang PY, Fan Y, Zhong Y, Dai Y, Mazloom AR, Chen EY, D'Agati V, Xiong H, Ross MJ, Chen N, Ma'ayan A, He JC
    Nat. Med., 2012-03-11;18(4):580-8.
    Species: Human
    Sample Types: Cell Lysates
  24. Therapeutic effect of Aralia cordata extracts on cartilage protection in collagenase-induced inflammatory arthritis rabbit model.
    Authors: Park DS, Huh JE, Baek YH
    J Ethnopharmacol, 2009-07-18;125(2):207-17.
    Species: Rabbit
    Sample Types: Serum
  25. Bakuchiol-induced caspase-3-dependent apoptosis occurs through c-Jun NH2-terminal kinase-mediated mitochondrial translocation of Bax in rat liver myofibroblasts.
    Authors: Park EJ, Zhao YZ, Kim YC, Sohn DH
    Eur. J. Pharmacol., 2007-01-23;559(2):115-23.
    Species: Mouse
    Sample Types: Cell Lysates
  26. Acylated and unacylated ghrelin promote proliferation and inhibit apoptosis of pancreatic beta-cells and human islets: involvement of 3',5'-cyclic adenosine monophosphate/protein kinase A, extracellular signal-regulated kinase 1/2, and phosphatidyl inositol 3-Kinase/Akt signaling.
    Authors: Granata R, Settanni F, Biancone L, Trovato L, Nano R, Bertuzzi F, Destefanis S, Annunziata M, Martinetti M, Catapano F, Ghè C, Isgaard J, Papotti M, Ghigo E, Muccioli G
    Endocrinology, 2006-10-26;148(2):512-29.
    Species: Hamster
    Sample Types: Cell Lysates
  27. Low dose leflunomide activates PI3K/Akt signalling in erythroleukemia cells and reduces apoptosis induced by anticancer agents.
    Authors: Leger DY, Liagre B, Beneytout JL
    Apoptosis, 2006-10-01;11(10):1747-60.
    Species: Human
    Sample Types: Cell Lysates

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