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Human VWF(Von Willebrand Factor) ELISA Kit

Citations(8)Uniprot : P04275

Cat.No.:E-EL-H2168
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	1.56-100 ng/mL
Sensitivity	0.94 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human VWF in samples. No significant cross-reactivity or interference between Human VWF 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 VWF 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 VWF. 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 VWF 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 VWF, 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 VWF. You can calculate the concentration of Human VWF 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 VWF were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human VWF 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)	4.94	13.78	40.52	4.74	13.70	42.12
Standard deviation	0.30	0.64	1.94	0.25	0.68	1.44
CV (%)	6.07	4.64	4.79	5.27	4.96	3.42

Recovery

The recovery of Human VWF 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)	90-106	97
EDTA plasma (n=8)	91-103	98
Cell culture media (n=8)	92-104	98

Linearity

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

Target Information

Database Links	SwissProt: P04275
Synonyms	F8VWF, VWD
Research Area	Cancer, Cardiovascular, Stem cells

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

THROMBOSIS RESEARCH (2022) IF: 10.407
High mobility group box 1 derived mainly from platelet microparticles exacerbates microvascular obstruction in no reflow

DOI: 10.1016/j.thromres.2022.12.003

Sample: Human cardiac microvascular endothelial cells (HCM
Antioxidants (2021) IF: 6.313
Resveratrol-Elicited PKC Inhibition Counteracts NOX-Mediated Endothelial to Mesenchymal Transition in Human Retinal Endothelial Cells Exposed to High Glucose

DOI: 10.3390/antiox10020224

PMID: 33540918
Sample: Cell lysate
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2019) IF: 4.183
Effect of Autophagy Regulated by Sirt1/FoxO1 Pathway on the Release of Factors Promoting Thrombosis from Vascular Endothelial Cells

DOI: 10.3390/ijms20174132

Sample: HUVEC cells
DIGESTIVE DISEASES AND SCIENCES (2022) IF: 3.487
Association of Heparin-Like Effect, Factor VII/XIII Deficiency and Fibrinolysis with Rebleeding Risk in Cirrhosis with Acute Variceal Bleeding

DOI: 10.1007/s10620-022-07656-9

Sample: serum
JOURNAL OF INTENSIVE CARE MEDICINE (2021) IF: 3.51
Prognostic Significance of Plasma Hepatocyte Growth Factor in Sepsis:

DOI: 10.1177/0885066621993423

Sample: Plasma
Annals of Translational Medicine (2021) IF: 3.932
Metformin alleviates bevacizumab-induced vascular endothelial injury by up-regulating GDF15 and activating the PI3K/AKT/FOXO/PPARγ signaling pathway

DOI: 10.21037/atm-21-4764

PMID: 34790753
Sample: HUVEC
World Journal of Clinical Cases (2020) IF: 1.013
Serum von Willebrand factor for early diagnosis of lung adenocarcinoma in patients with type 2 diabetes mellitus

DOI: 10.12998/wjcc.v8.i10.1916

PMID: 32518781
Sample: Serum
Brazilian Journal of Cardiovascular Surgery (2020) IF: 1.053
Comparison between the Effects of Bretschneider’s HTK Solution and Cold Blood Cardioplegia on Systemic Endothelial Functions in Patients who Undergo Coronary Artery Bypass Surgery: a Prospective Randomized and Controlled Trial

DOI: 10.21470/1678-9741-2019-0327

Sample: Serum

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