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Human COL4(Collagen Type Ⅳ) ELISA Kit

Cat.No.:E-EL-H0178
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.78-50 ng/mL
Sensitivity	0.47 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human COL4 in samples. No significant cross-reactivity or interference between Human COL4 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 COL4 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 COL4. 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 COL4 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 COL4, 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 COL4. You can calculate the concentration of Human COL4 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 COL4 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human COL4 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)	2.47	4.27	17.11	2.66	4.21	18.63
Standard deviation	0.16	0.23	0.53	0.17	0.19	0.87
CV (%)	6.48	5.39	3.10	6.39	4.51	4.67

Recovery

The recovery of Human COL4 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)	88-100	93
EDTA plasma (n=8)	92-106	99
Cell culture media (n=8)	86-102	93

Linearity

Samples were spiked with high concentrations of Human COL4 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Synonyms	Type-IV Collagen
Research Area	Cardiovascular, Signal transduction

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

CELL AND TISSUE RESEARCH (2022) IF: 4.051
Sestrin2 attenuates renal damage by regulating Hippo pathway in diabetic nephropathy

DOI: 10.1007/s00441-022-03668-z

PMID: 35821438
Sample: HMCs
PHYTOTHERAPY RESEARCH (2020) IF: 4.087
Ampelopsin inhibits high glucose‐induced extracellular matrix accumulation and oxidative stress in mesangial cells through activating the Nrf2/HO‐1 pathway

DOI: 10.1002/ptr.6668

Sample: Cell culture supernatant
Experimental and Therapeutic Medicine (2021) IF: 2.447
Knockout of NGAL aggravates tubulointerstitial injury in a mouse model of diabetic nephropathy by enhancing oxidative stress and fibrosis

DOI: 10.3892/etm.2021.9752

PMID: 33732294
Sample: Cell culture supernatant
EUROPEAN JOURNAL OF GASTROENTEROLOGY & HEPATOLOGY (2021) IF: 2.568
Serum fibrosis markers could aid in the prediction of factor for successful oral antiviral treatment in hepatitis C

DOI: 10.1097/MEG.0000000000002083

PMID: 33560684
Sample: Blood
Journal of Diabetes Research (2020) IF: 2.965
Gremlin in the Vitreous of Patients with Proliferative Diabetic Retinopathy and the Downregulation of Gremlin in Retinal Pigment Epithelial Cells

DOI: 10.1155/2020/9238742

PMID: 32377526
Sample: Cell culture supernatant,Tissue homogenate

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