ELISA Kit for S100 Calcium Binding Protein A6 (S100A6) Rattus norvegicus (Rat) Sandwich ELISA

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

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  • ELISA Kit for S100 Calcium Binding Protein A6 (S100A6) Packages (Simulation)
  • ELISA Kit for S100 Calcium Binding Protein A6 (S100A6) Packages (Simulation)
  • ELISA Kit for S100 Calcium Binding Protein A6 (S100A6) Results demonstration
  • SEB769Ra.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 A6 (S100A6) and the recovery rates were calculated by comparing the measured value to the expected amount of S100 Calcium Binding Protein A6 (S100A6) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 94-103 101
EDTA plasma(n=5) 82-104 96
heparin plasma(n=5) 86-97 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level S100 Calcium Binding Protein A6 (S100A6) 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) 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) 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) 81-101% 89-96% 90-97% 79-89%
EDTA plasma(n=5) 89-98% 85-93% 95-102% 86-97%
heparin plasma(n=5) 81-102% 88-103% 82-90% 79-102%

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 A6 (S100A6)

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 A6 (S100A6). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to S100 Calcium Binding Protein A6 (S100A6). 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 A6 (S100A6), 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 A6 (S100A6) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Tear proteome and protein network analyses reveal a novel pentamarker panel for tear film characterization in dry eye and meibomian gland dysfunctionPubMed: 23201116
  • Diagnostic significance of S100A2 and S100A6 levels in sera of patients with non-small cell lung cancerPubMed: 26361956
  • Serum levels of S100A6 are unaltered in patients with resectable cholangiocarcinomapubmed:27709523
  • 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
  • Proteomics detection of S100A6 in tumor tissue interstitial fluid and evaluation of its potential as a biomarker of cholangiocarcinomaPubmed:29629840
  • Serum S100A6, S100A8, S100A9 and S100A11 proteins in colorectal neoplasia: results of a single centre prospective studyPubmed: 31856598
  • S100A6 represses Calu©\6 lung cancer cells growth via inhibiting cell proliferation, migration, invasion and enhancing apoptosis34008212
  • Quantification of a panel for dry-eye protein biomarkers in tears: A comparative pilot study using standard ELISA and customized microarrays34012227

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