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Human S100A12(S100 Calcium Binding Protein A12) ELISA Kit

Citations(5)Uniprot : P80511

Cat.No.:E-EL-H1293
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 S100A12 in samples. No significant cross-reactivity or interference between Human S100A12 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 S100A12 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 S100A12. 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 S100A12 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 S100A12, 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 S100A12. You can calculate the concentration of Human S100A12 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 S100A12 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human S100A12 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)	0.49	1.08	4.12	0.48	1.06	3.86
Standard deviation	0.03	0.05	0.14	0.03	0.05	0.15
CV (%)	6.12	4.63	3.40	6.25	4.72	3.89

Recovery

The recovery of Human S100A12 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)	93-107	98
EDTA plasma (n=8)	91-107	98
Cell culture media (n=8)	89-102	95

Linearity

Samples were spiked with high concentrations of Human S100A12 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P80511
Synonyms	CAAF1, CAGC, CGRP, ENRAGE, MRP-6, MRP6, p6, Calgranulin C
Research Area	Cancer, Immunology, 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

CLINICAL AND EXPERIMENTAL IMMUNOLOGY (2022) IF: 4.33
Clinical features and nasal inflammation in asthma and allergic rhinitis

DOI: 10.1093/cei/uxac019

Sample: blood
CELLULAR ONCOLOGY (2018) IF: 4.761
TLR4 and RAGE conversely mediate pro-inflammatory S100A8/9-mediated inhibition of proliferation-linked signaling in myeloproliferative neoplasms

DOI: 10.1007/s13402-018-0392-6

Sample: Plasma
Journal of Clinical Lipidology (2018) IF: 3.58
Identification of proresolving and inflammatory lipid mediators in human psoriasis

DOI: 10.1016/j.jacl.2018.03.091

Sample: serum
Biomarkers in Medicine (2017) IF: 2.02
Enhanced S100A9 and S100A12 expression in acute coronary syndrome

DOI: 10.2217/bmm-2016-0253

Sample: Serum
International Journal of Rheumatic Diseases (2022) IF: 2.558
The role of S100A12 and Toll-like receptor 4 in assessment of disease activity in familial Mediterranean fever and juvenile idiopathic arthritis

DOI: 10.1111/1756-185X.14475

PMID: 36302511
Sample: blood

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