ELISA Kit for S100 Calcium Binding Protein A8 (S100A8) Sus scrofa; Porcine (Pig) Sandwich ELISA

MRP8; MIF; 60B8AG; CAGA; CFAG; CGLA; CP-10; L1Ag; NIF; P8; Calgranulin A; Cystic fibrosis antigen; Leukocyte L1 complex light chain; Migration inhibitory factor-related protein 8

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  • ELISA Kit for S100 Calcium Binding Protein A8 (S100A8) Packages (Simulation)
  • ELISA Kit for S100 Calcium Binding Protein A8 (S100A8) Packages (Simulation)
  • ELISA Kit for S100 Calcium Binding Protein A8 (S100A8) Results demonstration
  • SEB792Po.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant S100 Calcium Binding Protein A8 (S100A8) and the recovery rates were calculated by comparing the measured value to the expected amount of S100 Calcium Binding Protein A8 (S100A8) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-102 95
EDTA plasma(n=5) 90-97 93
heparin 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 A8 (S100A8) 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 A8 (S100A8) 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 A8 (S100A8) 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) 90-104% 88-96% 91-99% 80-98%
EDTA plasma(n=5) 95-104% 78-99% 96-105% 88-102%
heparin plasma(n=5) 85-105% 80-98% 93-105% 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
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.

ELISA Kit for S100 Calcium Binding Protein A8 (S100A8)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to S100 Calcium Binding Protein A8 (S100A8). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to S100 Calcium Binding Protein A8 (S100A8). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain S100 Calcium Binding Protein A8 (S100A8), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of S100 Calcium Binding Protein A8 (S100A8) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • 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 miceScienceDirect: S0006291X1200352X
  • Association of Increased S100A8 Serum Protein with Early Pregnancy LossPubmed:25252120
  • Soluble receptor for advanced glycation end products inhibits disease progression in autosomal dominant polycystic kidney disease by down-regulating cell proliferationPubMed: 25934702
  • S100A8 Production in CXCR2-Expressing CD11b+ Gr-1high Cells Aggravates Hepatitis in Mice Fed a High-Fat and High-Cholesterol DietPubMed: 26608915
  • Analytical Performance of ELISA Assays in Urine: One More Bottleneck towards Biomarker Validation and Clinical ImplementationPubmed:26889680
  • Proteomics Analysis of Blood Serums from Alzheimer's Disease Patients Using iTRAQ LabelingTechnology.pubmed:27911324
  • Standardized ethanolic extract of the rhizome of Curcuma xanthorrhiza prevents murine ulcerative colitis by regulation of inflammationS1756464617300208
  • Proinflammatory effects of S100A8/A9 via TLR4 and RAGE signaling pathways in BV-2 microglial cellspubmed:28498464
  • Immune Modulating Topical S100A8/A9 Inhibits Growth of Pseudomonas aeruginosa and Mitigates Biofilm Infection in Chronic Wounds.pubmed:28672877
  • Tu1938-Micronutrients Assessment in Patients with Cirrhosis10.1016:S0016-5085(18)33544-3
  • Tu1942-PI3K/AKT-Mediated Upregulation of C7 Inhibits Colorectal Cancer Proliferation and Metastasis10.1016:S0016-5085(18)33544-5
  • Signaling between pancreatic β-cells and macrophages via S100 calcium-binding protein A8 exacerbates β-cell apoptosis and islet inflammationPubmed:29496993
  • S100A8/A9 promotes MMP-9 expression in the fibroblasts from cardiac rupture after myocardial infarction by inducing macrophages secreting TNFα.Pubmed:29949169
  • Synergistic effect of immunomodulatory S100A8/A9 and ciprofloxacin against Pseudomonas aeruginosa biofilm in a murine chronic wound modelPubmed: 31116394
  • Systemic Inflammation Precedes Microalbuminuria in Diabetes
  • iTRAQ-based quantitative protein expression profiling of biomarkers in childhood B-cell and T-cell acute lymphoblastic leukemiaPubmed: 31440093
  • Serum S100A6, S100A8, S100A9 and S100A11 proteins in colorectal neoplasia: results of a single centre prospective studyPubmed: 31856598
  • Analysis of novel serum markers of fibrosis and angiogenesis in patients with alcoholic liver cirrhosis
  • Quantification of a panel for dry-eye protein biomarkers in tears: A comparative pilot study using standard ELISA and customized microarrays34012227
  • Adjunctive S100A8/A9 Immunomodulation Hinders Cipro fl oxacin Resistance in

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