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Human Cys-C(Cystatin C) ELISA Kit

Cat.No.:E-EL-H3643
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 Cys-C in samples. No significant cross-reactivity or interference between Human Cys-C 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 Cys-C 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 Cys-C. 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 Cys-C 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 Cys-C, 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 Cys-C. You can calculate the concentration of Human Cys-C 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 Cys-C were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human Cys-C 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)	2.30	6.80	15.60	2.30	7.50	15.60
Standard deviation	0.09	0.12	0.23	0.10	0.16	0.24
CV (%)	4.55	4.22	5.28	6.13	5.73	4.29

Recovery

The recovery of Human Cys-C 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)	90-105	98
EDTA plasma (n=8)	94-110	100
Cell culture media (n=8)	92-101	97

Linearity

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

Target Information

Synonyms	CST3, Cystatin 3, ARMD11, Post-Gamma-Globulin
Research Area	Cancer, Cell Biology, Cardiovascular, Stem cells, Tags and Cell Markers

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

Citations

Translational Research (2022) IF: 7.012
Interplay between mineral bone disorder and cardiac damage in acute kidney injury: from Ca2+ mishandling and preventive role of Klotho in mice to its potential mortality prediction in human

DOI: 10.1016/j.trsl.2022.01.002

PMID: 35077866
Sample: plasma
Journal of Inflammation Research (2021) IF: 6.922
Endothelin-1/Endothelin Receptor Type A-Angiopoietins/Tie-2 Pathway in Regulating the Cross Talk Between Glomerular Endothelial Cells and Podocytes in Trichloroethylene-Induced Renal Immune Injury

DOI: 10.2147/JIR.S301104

Sample: Serum,Plasma,Urine
Nutrients (2018) IF: 4.196
Tocotrienol-Rich Vitamin E from Palm Oil (Tocovid) and Its Effects in Diabetes and Diabetic Nephropathy: A Pilot Phase II Clinical Trial

DOI: 10.3390/nu10091315

PMID: 30227659
Sample: Serum
JOURNAL OF NEPHROLOGY (2023) IF: 4.393
Associations of prematurity and low birth weight with blood pressure and kidney function in middle-aged participants of the Brazilian Longitudinal Study of Adult Health: ELSA-Brasil

DOI: 10.1007/s40620-022-01549-w

Sample: Serum
PLoS One (2021) IF: 3.24
Effects of pneumoperitoneum on kidney injury biomarkers: A randomized clinical trial

DOI: 10.1371/journal.pone.0247088

PMID: 33606739
Sample: Plasma
Biomed Research International (2022) IF: 3.246
Evaluation Value of Serum miR-4299 and miR-16-5p in Risk Stratification of Sepsis-Induced Acute Kidney Injury

DOI: 10.1155/2022/5165892

PMID: 35845963
Sample: serum
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2016) IF: 3.257
Label-Free Quantitative Proteomics Reveals Differences in Molecular Mechanism of Atherosclerosis Related and Non-Related to Chronic Kidney Disease

DOI: 10.3390/ijms17050631

Sample: Plasma
MEDICINE (2018) IF: 2.028
Long-term benefits of high-intensity atorvastatin therapy in Chinese acute coronary syndrome patients undergoing percutaneous coronary intervention: A retrospective study

DOI: 10.1097/MD.0000000000012687

PMID: 30334951
Sample: Serum
SAUDI MEDICAL JOURNAL (2020) IF: 1.195
Evaluation of early biomarkers of renal dysfunction in diabetic patients.

DOI: 10.15537/smj.2020.7.25168

Sample: serum
Genetic Testing and Molecular Biomarkers (2020) IF: 1.378
Correlation Between Single Nucleotide Polymorphisms at the 3′-UTR of the NFKB1 Gene and Acute Kidney Injury in Sepsis

DOI: 10.1089/gtmb.2019.0222

Sample: Cell sample

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