ELISA Kit for S100 Calcium Binding Protein A9 (S100A9)

MRP14; MIF; 60B8AG; CFAG; NIF; CAGB; CGLB; L1AG; P14; Calgranulin B; Migration inhibitory factor-related protein 14; Calprotectin L1H subunit; Leukocyte L1 complex heavy chain

Specificity

This assay has high sensitivity and excellent specificity for detection of S100 Calcium Binding Protein A9 (S100A9).
No significant cross-reactivity or interference between S100 Calcium Binding Protein A9 (S100A9) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant S100 Calcium Binding Protein A9 (S100A9) and the recovery rates were calculated by comparing the measured value to the expected amount of S100 Calcium Binding Protein A9 (S100A9) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-105 101
EDTA plasma(n=5) 95-104 101
heparin plasma(n=5) 83-105 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level S100 Calcium Binding Protein A9 (S100A9) 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 A9 (S100A9) 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 A9 (S100A9) 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) 95-102% 89-103% 79-96% 79-103%
EDTA plasma(n=5) 81-92% 95-102% 78-93% 90-98%
heparin plasma(n=5) 99-105% 93-101% 80-103% 80-97%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
J. Proteome Res. Differential Proteomic Analysis Distinguishes Tissue Repair Biomarker Signatures in Wound Exudates Obtained from Normal Healing and Chronic Wounds Pubmed: source
Biochemical and Biophysical Research Communications High circulating levels of S100A8/A9 complex (calprotectin) in male Japanese with abdominal adiposity and dysregulated expression of S100A8 and S100A9 in adipose tissues of obese mice ScienceDirect: S0006291X1200352X
PLoS One. Proteins with Altered Levels in Plasma from Glioblastoma Patients as Revealed by iTRAQ-Based Quantitative Proteomic Analysis PubMed: 23029420
Mainz Identifizierung neuer potentieller Biomarker für das Kolonkarzinom sowie dessen Vorstufen Info:Source
PLOS ONE Susceptibility to COPD: Differential Proteomic Profiling after Acute Smoking Pubmed:25036363
the faseb journal Soluble receptor for advanced glycation end products inhibits disease progression in autosomal dominant polycystic kidney disease by down-regulating cell proliferation PubMed: 25934702
Sci Rep.  S100A9 and ORM1 serve as predictors of therapeutic response and prognostic factors in advanced extranodal NK/T cell lymphoma patients treated with pegaspargase/gemcitabine pmc:PMC4810364
Journal of Alzheimer's Disease Proteomics Analysis of Blood Serums from Alzheimer's Disease Patients Using iTRAQ LabelingTechnology. pubmed:27911324
International journal of molecular sciences Immune Modulating Topical S100A8/A9 Inhibits Growth of Pseudomonas aeruginosa and Mitigates Biofilm Infection in Chronic Wounds. pubmed:28672877
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
Journal of Biological Chemistry Signaling between pancreatic β-cells and macrophages via S100 calcium-binding protein A8 exacerbates β-cell apoptosis and islet inflammation Pubmed:29496993
Cancer Management and Research The role of calgranulin B gene on the biological behavior of squamous cervical cancer in vitro and in vivo Pubmed:29497331
European review for medical and pharmacological sciences S100A8/A9 promotes MMP-9 expression in the fibroblasts from cardiac rupture after myocardial infarction by inducing macrophages secreting TNFα. Pubmed:29949169
Estudio de células inmunosupresoras y biomarcadores circulantes en carcinoma renal
Lung Mass Spectrometry Analysis of the Exhaled Breath Condensate and Proposal of Dermcidin and S100A9 as Possible Markers for Lung Cancer Prognosis Pubmed: 31115649
中国医药导报 降钙素原, 超敏C 反应蛋白, 红细胞沉降率及S100A9 对肺结核合并肺部细菌感染的影响
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
PLoS One Adverse reproductive effects of S100A9 on bovine sperm and early embryonic development in vitro Pubmed: 31945120
Sci Rep Overexpression of S100A9 in tumor stroma contribute to immune evasion of NK/T cell lymphoma and predict poor response rate 34045609
university of copenhagen Adjunctive S100A8/A9 Immunomodulation Hinders Cipro fl oxacin Resistance in
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