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Mouse β-CTx(Beta Crosslaps) ELISA Kit

Cat.No.:E-EL-M0372
Reactivity: Mouse

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96T
48T
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96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	78.13-5000 pg/mL
Sensitivity	46.88 pg/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes β-CTx in samples. No significant cross-reactivity or interference between β-CTx 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 β-CTx concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with β-CTx. During the reaction, β-CTx in the sample or standard competes with a fixed amount of β-CTx on the solid phase supporter for sites on the Biotinylated Detection Ab specific to β-CTx. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of β-CTx in tested samples can be calculated 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℃
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 β-CTx were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level β-CTx 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 (pg/mL)	264.20	768.90	1936.80	252.10	764.90	1935.40
Standard deviation	16.10	34.60	93.00	14.60	32.90	71.60
CV (%)	6.09	4.50	4.80	5.79	4.30	3.70

Recovery

The recovery of β-CTx 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)	88-103	94
EDTA plasma(n=8)	91-104	98
Cell culture media(n=8)	85-96	91

Linearity

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

Target Information

Synonyms	b-CTx, bCTXI, bCTX-I, B-Cr, BCL, Type I Collagen C-Telopeptide-Related Fraction
Research Area	Signal transduction, Metabolism

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

Journal of the American Chemical Society (2020) IF: 14.612
Smart Nanosacrificial Layer on the Bone Surface Prevents Osteoporosis through Acid–Base Neutralization Regulated Biocascade Effects

DOI: 10.1021/jacs.0c07309

PMID: 32960592
Sample: Serum
BIOMEDICINE & PHARMACOTHERAPY (2022) IF: 7.419
Thiaplakortone B attenuates RANKL‐induced NF‐κB and MAPK signaling and dampens OVX‐induced bone loss in mice

DOI: 10.1016/j.biopha.2022.113622

Sample: serum
Cell Death & Disease (2020) IF: 6.304
PKC-δ deficiency in B cells displays osteopenia accompanied with upregulation of RANKL expression and osteoclast–osteoblast uncoupling

DOI: 10.1038/s41419-020-02947-3

Sample: Serum
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE (2014) IF: 5.089
Discoidin domain receptor 2 facilitates prostate cancer bone metastasis via regulating parathyroid hormone-related protein

DOI: 10.1016/j.bbadis.2014.04.018

Sample: Cell culture supernatant
INTERNATIONAL IMMUNOPHARMACOLOGY (2023) IF: 5.714
Cedrol, a Ginger-derived sesquiterpineol, suppresses estrogen-deficient osteoporosis by intervening NFATc1 and reactive oxygen species

DOI: 10.1016/j.intimp.2023.109893

Sample: serum
JOURNAL OF BONE AND MINERAL RESEARCH (2020) IF: 5.854
Hymenialdisine: A Marine Natural Product That Acts on Both Osteoblasts and Osteoclasts and Prevents Estrogen-Dependent Bone Loss in Mice

DOI: 10.1002/jbmr.4025

Sample: serum

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