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Human CASP3(Caspase 3) ELISA Kit

Citations(12)Uniprot : P42574

Cat.No.:E-EL-H0017
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human CASP3 in samples. No significant cross-reactivity or interference between Human CASP3 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human CASP3 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human CASP3. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human CASP3 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human CASP3, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human CASP3. You can calculate the concentration of Human CASP3 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CASP3 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CASP3 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	0.97	2.79	7.18	0.94	2.57	7.67
Standard deviation	0.05	0.12	0.32	0.06	0.14	0.27
CV (%)	5.15	4.30	4.46	6.38	5.45	3.52

Recovery

The recovery of Human CASP3 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	87-100	93
EDTA plasma (n=8)	88-103	94
Cell culture media (n=8)	93-106	99

Linearity

Samples were spiked with high concentrations of Human CASP3 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P42574
Synonyms	CPP32, CPP32B, SCA-1, Apoptain, Yama
Research Area	Cancer, Cell Biology, Metabolism, Neuroscience

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

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Citations

Antioxidants (2022) IF: 6.313
Oxidant/Antioxidant Status Is Impaired in Sepsis and Is Related to Anti-Apoptotic, Inflammatory, and Innate Immunity Alterations

DOI: 10.3390/antiox11020231

PMID: 35204114
Sample: serum
Pharmaceutics (2021) IF: 6.321
Comparative Study of the Pharmacological Properties and Biological Effects of Polygonum aviculare L. herba Extract-Entrapped Liposomes versus Quercetin-Entrapped Liposomes on Doxorubicin-Induced Toxicity on HUVECs

DOI: 10.3390/pharmaceutics13091418

PMID: 34575493
Sample: HUVEC
PHYTOMEDICINE (2019) IF: 4.18
Ethyl acetate extract of Caesalpinia sappan L. inhibited acute myeloid leukemia via ROS-mediated apoptosis and differentiation

DOI: 10.1016/j.phymed.2019.153142

PMID: 32045840
Sample: Cell lysate
Scientific Reports (2021) IF: 4.38
Survivin and caspases serum protein levels and survivin variants mRNA expression in sepsis

DOI: 10.1038/s41598-020-78208-2

PMID: 33441606
Sample: Serum
INTERNATIONAL JOURNAL OF CARDIOLOGY (2016) IF: 4.638
Melatonin effects on myocardial ischemia–reperfusion injury: Impact on the outcome in patients undergoing coronary artery bypass grafting surgery

DOI: 10.1016/j.ijcard.2016.07.108

Sample: Plasma
Reproductive Biology and Endocrinology (2020) IF: 3.235
Growth hormone activates PI3K/Akt signaling and inhibits ROS accumulation and apoptosis in granulosa cells of patients with polycystic ovary syndrome

DOI: 10.1186/s12958-020-00677-x

PMID: 33287836
Sample: Cell culture supernatant
MOLECULES (2020) IF: 3.267
Development of HPLC Method for Quantification of Sinigrin from Raphanus sativus Roots and Evaluation of Its Anticancer Potential

DOI: 10.3390/molecules25214947

PMID: 33114598
Sample: Cell culture supernatant
NANOTECHNOLOGY (2020) IF: 3.551
Targeted gold nanorods combined with low-intensity nsPEFs enhance antimelanoma efficacy in vitro

DOI: 10.1088/1361-6528/ab925f

Sample: A375 cell
ONCOLOGY REPORTS (2021) IF: 3.906
BET inhibitors combined with chemotherapy synergistically inhibit the growth of NSCLC cells

DOI: 10.3892/or.2021.8021

PMID: 33760217
Sample: A549 cell,H157 cell
BIOMARKERS (2017) IF: 2.006
Circulating levels of apoptotic markers and oxidative stress parameters in women with polycystic ovary syndrome: a case-controlled descriptive study

DOI: 10.1080/1354750X.2016.1265004

Sample: Serum
Journal of Nanomaterials (2021) IF: 2.986
High Expressions of Notch and Survivin in Elderly Patients with Glioma Contribute to an Unfavorable Prognosis

DOI: 10.1155/2021/9942260

Sample: Serum
GYNECOLOGIC AND OBSTETRIC INVESTIGATION (2018) IF: 1.183
The Role of Serum Caspase 3 Levels in Prediction of Endometriosis Severity

DOI: 10.1159/000489494

Sample: Serum

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