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Human NTXⅠ(Cross Linked N-telopeptide of Type Ⅰ Collagen) ELISA Kit

Citations(4)

Cat.No.:E-EL-H0836
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T/24
Assay time	3.5h
Detection range	3.13-200 ng/mL
Sensitivity	1.88 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human NTXⅠ in samples. No significant cross-reactivity or interference between Human NTXⅠ 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 NTXⅠ 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 NTXⅠ. 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 NTXⅠ 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 NTXⅠ, 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 NTXⅠ. You can calculate the concentration of Human NTXⅠ 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℃, 12 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), 12 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 12 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 NTXⅠ were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human NTXⅠ 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)	9.27	30.79	98.10	8.47	30.12	97.13
Standard deviation	0.54	1.35	3.18	0.43	1.50	2.93
CV (%)	5.83	4.38	3.24	5.08	4.98	3.02

Recovery

The recovery of Human NTXⅠ 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)	86-98	91
EDTA plasma (n=8)	87-102	93
Cell culture media (n=8)	91-103	98

Linearity

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

Target Information

Synonyms	NTX-I, NTX1,NTX
Research Area	Metabolism, Cancer

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

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2021) IF: 5.924

3D Environment Is Required In Vitro to Demonstrate Altered Bone Metabolism Characteristic for Type 2 Diabetics

DOI: 10.3390/ijms22062925
PMID: 33805833	Sample: Cell Culture Supernatant(Thp-1 Cells)

OSTEOARTHRITIS AND CARTILAGE (2017) IF: 4.742

Associations between serum ghrelin and knee symptoms, joint structures and cartilage or bone biomarkers in patients with knee osteoarthritis

DOI: 10.1016/j.joca.2017.05.015
	Sample: Serum

Bosnian Journal of Basic Medical Sciences (2018) IF: 1.432

Evaluation of bone mineral density (BMD) and indicators of bone turnover in patients with hemophilia

DOI: 10.17305/bjbms.2018.2335
	Sample: Urine

CLINICAL RHEUMATOLOGY (2019) IF: 2.293

Associations between serum IL-8 and knee symptoms, joint structures, and cartilage or bone biomarkers in patients with knee osteoarthritis

DOI: 10.1007/s10067-019-04718-8
PMID: 31377918	Sample: Serum

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Reviews/Q&A

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Reviews
Q&A

K***********aSubmitted [ Apr 10 2024 ]

Asked: Inquiry Regarding EDTA Plasma NTX Measurement I am writing to inquire about the measurement of NTX in human EDTA plasma using the ELISA kit E-EL-H0836, Lot No. WX0840484776, which we recently purchased from your company. Upon conducting measurements, we observed that the standard samples worked properly; however, the measurement values for 100uL of EDTA plasma indicated levels below the detection limit of 3.13 ng/mL. Could you please provide information on the expected range of NTX measurements in human EDTA plasma using this specific ELISA kit? Additionally, any recommendations for optimizing the assay in EDTA plasma samples would be greatly appreciated. Thank you for your attention to this matter. I look forward to your prompt response. Sincerely, Kotaro Tamura, Ph.D Kao Corporation, Japan

adminSubmitted [ Apr 10 2024 ]

Answered: Hello Tamura-san,we have validated E-EL-H0836 with healthy human plasma, all the samples were undiluted and the reference value was ND-25 ng/mL.

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