Multiplex Assay Kit for S100 Calcium Binding Protein A12 (S100A12) ,etc. by FLIA (Flow Luminescence Immunoassay)

CGRP; P6; CAAF1; CAGC; ENRAGE; MRP6; Calgranulin C; Calcium-binding protein in amniotic fluid 1; Extracellular newly identified RAGE-binding; Migration inhibitory factor-related 6

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of S100 Calcium Binding Protein A12 (S100A12) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between S100 Calcium Binding Protein A12 (S100A12) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant S100 Calcium Binding Protein A12 (S100A12) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of S100 Calcium Binding Protein A12 (S100A12) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-90 80
EDTA plasma(n=5) 78-89 81
heparin plasma(n=5) 94-101 97
sodium citrate plasma(n=5) 83-99 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level S100 Calcium Binding Protein A12 (S100A12) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level S100 Calcium Binding Protein A12 (S100A12) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of S100 Calcium Binding Protein A12 (S100A12) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 88-95% 99-105% 80-95% 82-101%
EDTA plasma(n=5) 92-105% 79-98% 87-105% 94-102%
heparin plasma(n=5) 89-104% 86-94% 89-96% 84-104%
sodium citrate plasma(n=5) 88-101% 88-101% 85-103% 97-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:S100A12) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Inflammation Research Correlation of human S100A12 (EN-RAGE) and high-sensitivity C-reactive protein as gingival crevicular fluid and serum markers of inflammation in chronic periodontitis and type 2 diabetes Pubmed: 24378957
Dig Dis Sci The Circulating Level of Soluble Receptor for Advanced Glycation End Products Displays Different Patterns in Ulcerative Colitis and Crohn’s Disease: A Cross-Sectional Study PubMed: 25757448
The Journal of Clinical Endocrinology & Metabolism Low Endogenous Secretory Receptor for Advanced Glycation End-Products Levels Are Associated With Inflammation and Carotid Atherosclerosis in Prediabetes Pubmed:26885882
Nutrition, Metabolism, and Cardiovascular Diseases High intake of dietary advanced glycation end-products is associated with increased arterial stiffness and inflammation in subjects with type 2 diabetes. pubmed:28958695
The Journal of Clinical Endocrinology & Metabolism HbA1c Identifies Subjects With Prediabetes and Subclinical Left Ventricular Diastolic Dysfunction. pubmed:28973588
Journal of Immunology Research A Granulocyte-Specific Protein S100A12 as a Potential Prognostic Factor Affecting Aggressiveness of Therapy in Patients with Juvenile Idiopathic Arthritis Doi: 10.1155/2018/5349837
Al-Azhar Assiut Medical Journal The role of serum S100A12 (calgranulin C) as a diagnostic marker in Egyptian patients with irritable bowel syndrome and ulcerative colitis Doi: 10.4103/AZMJ.AZMJ_52_18
Acta Diabetologica Analysis of S100A12 plasma levels in hyperlipidemic subjects with or without familial hypercholesterolemia Pubmed: 30963307
Cells 1 h Postload Glycemia Is Associated with Low Endogenous Secretory Receptor for Advanced Glycation End Product Levels and Early Markers of Cardiovascular …
Biomolecules . Multimarker Approach to Identify Patients with Coronary Artery Disease at High Risk for Subsequent Cardiac Adverse Events: The Multi-Biomarker Study Pubmed: 32549327
Mediators of Inflammation Alarmins HMGB1, IL-33, S100A7, and S100A12 in Psoriasis Vulgaris
Animals (Basel) The Serum and Saliva Proteome of Dogs with Diabetes Mellitus 33271797
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