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Human CTXⅡ(Cross Linked C-telopeptide of Type Ⅱ Collagen) ELISA Kit

Cat.No.:E-EL-H0837
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T
Assay time	3.5h
Detection range	0.16-10 ng/mL
Sensitivity	0.10 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human CTXⅡ in samples. No significant cross-reactivity or interference between Human CTXⅡ 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 CTXⅡ 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 CTXⅡ. 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 CTXⅡ 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 CTXⅡ, 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 CTXⅡ. You can calculate the concentration of Human CTXⅡ 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 CTXⅡ were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CTXⅡ 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)	0.50	1.10	4.60	0.50	1.00	4.20
Standard deviation	0.03	0.05	0.14	0.03	0.06	0.20
CV (%)	6.00	4.55	3.04	6.00	6.00	4.76

Recovery

The recovery of Human CTXⅡ 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)	92-103	98
EDTA plasma (n=8)	88-98	93
Cell culture media (n=8)	85-100	92

Linearity

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

Target Information

Synonyms	CTX2
Research Area	Metabolism

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

BIOSENSORS & BIOELECTRONICS (2014) IF: 6.451
Detection of CTX-II in serum and urine to diagnose osteoarthritis by using a fluoro-microbeads guiding chip

DOI: 10.1016/j.bios.2014.08.016

Sample: Serum,Urine
Biomed Research International (2018) IF: 2.583
Clinical and Biological Signature of Osteochondritis Dissecans in a Cross-Sectional Study

DOI: 10.1155/2018/5458704

Sample: Serum
Biomed Research International (2018) IF: 2.583
Cartilage and Bone Serum Biomarkers as Novel Tools for Monitoring Knee Osteochondritis Dissecans Treated with Osteochondral Scaffold

DOI: 10.1155/2018/9275102

Sample: serum
Evidence-based Complementary and Alternative Medicine (2022) IF: 2.63
Tamarindus indica Seed Extract-Based Botanical Compositions Alleviate Knee Pain and Improve Joint Function in Mild-to-Moderate Osteoarthritis: A Randomized, Double-Blind, Placebo-Controlled Clinical Study

DOI: 10.1155/2022/2226139

PMID: 35096104
Sample: serum
Central European Journal of Immunology (2020) IF: 1.415
Joint remodeling outcome of serum levels of Dickkopf-1 (DKK1), cartilage oligomeric matrix protein (COMP), and C-telopeptide of type II collagen (CTXII) in rheumatoid arthritis

DOI: 10.5114/ceji.2020.94685

PMID: 32425683
Sample: Serum
BMC MUSCULOSKELETAL DISORDERS (2018) IF: 1.998
Influence of a periodized circuit training protocol on intermuscular adipose tissue of patients with knee osteoarthritis: protocol for a randomized controlled trial

DOI: 10.1186/s12891-018-2325-y

PMID: 30497420
Sample: Urine
Archives of Rheumatology (2016) IF: 0.22
Which Predicts Quadriceps Muscle Strength in Knee Osteoarthritis: Biological Markers or Clinical Variables?

DOI: 10.5606/ArchRheumatol.2017.5919

PMID: 30375528
Sample: Serum

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Reviews/Q&A

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Reviews
Q&A

K*****ySubmitted [ Aug 23 2022 ]

Asked: Kindly let me know the sample preparation prior to the assay analysis for urine

adminSubmitted [ Aug 23 2022 ]

Answered: Use a sterile container to collect urine samples. Remove particulates by centrifugation for 15 minutes at 1000×g at 2-9°C. Collect the supernatant to carry out the assay

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