Multiplex Assay Kit for S100 Calcium Binding Protein A6 (S100A6) ,etc. by FLIA (Flow Luminescence Immunoassay)

S100-A6; 2A9; 5B10; CABP; CACY; PRA; MLN 4; Calcyclin; Growth factor-inducible protein 2A9; Prolactin receptor-associated protein

(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 A6 (S100A6) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between S100 Calcium Binding Protein A6 (S100A6) ,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 A6 (S100A6) ,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 A6 (S100A6) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-102 94
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 85-93 88
sodium citrate plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level S100 Calcium Binding Protein A6 (S100A6) ,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 A6 (S100A6) ,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 A6 (S100A6) ,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) 86-101% 87-94% 78-104% 81-101%
EDTA plasma(n=5) 88-101% 79-96% 91-98% 84-101%
heparin plasma(n=5) 88-99% 89-102% 79-103% 83-93%
sodium citrate plasma(n=5) 92-99% 91-104% 93-101% 96-105%

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#:S100A6) 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
Journal of Proteomics Tear proteome and protein network analyses reveal a novel pentamarker panel for tear film characterization in dry eye and meibomian gland dysfunction PubMed: 23201116
Tumour Biol Diagnostic significance of S100A2 and S100A6 levels in sera of patients with non-small cell lung cancer PubMed: 26361956
Clinical and Translational Medicine Serum levels of S100A6 are unaltered in patients with resectable cholangiocarcinoma pubmed:27709523
Gastroenterology Tu1938-Micronutrients Assessment in Patients with Cirrhosis 10.1016:S0016-5085(18)33544-3
Gastroenterology Tu1942-PI3K/AKT-Mediated Upregulation of C7 Inhibits Colorectal Cancer Proliferation and Metastasis 10.1016:S0016-5085(18)33544-5
Tumor Biology Proteomics detection of S100A6 in tumor tissue interstitial fluid and evaluation of its potential as a biomarker of cholangiocarcinoma Pubmed:29629840
scandinavian journal of clinical & laboratory investigation Serum S100A6, S100A8, S100A9 and S100A11 proteins in colorectal neoplasia: results of a single centre prospective study Pubmed: 31856598
CELL BIOCHEMISTRY AND FUNCTION S100A6 represses Calu©\6 lung cancer cells growth via inhibiting cell proliferation, migration, invasion and enhancing apoptosis 34008212
MOLECULAR VISION Quantification of a panel for dry-eye protein biomarkers in tears: A comparative pilot study using standard ELISA and customized microarrays 34012227
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