ELISA Kit for Anterior Gradient Protein 2 (AGR2) Homo sapiens (Human) Sandwich ELISA

GOB4; HAG2; XAG2; AG2; PDIA17; HPC8; Protein Disulfide Isomerase Family A,Member 17; Secreted cement gland protein XAG-2 homolog

  • ELISA Kit for Anterior Gradient Protein 2 (AGR2) Packages (Simulation)
  • ELISA Kit for Anterior Gradient Protein 2 (AGR2) Packages (Simulation)
  • ELISA Kit for Anterior Gradient Protein 2 (AGR2) Results demonstration
  • SEC285Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Anterior Gradient Protein 2 (AGR2) and the recovery rates were calculated by comparing the measured value to the expected amount of Anterior Gradient Protein 2 (AGR2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-96 89
EDTA plasma(n=5) 86-102 98
heparin plasma(n=5) 86-95 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anterior Gradient Protein 2 (AGR2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Anterior Gradient Protein 2 (AGR2) 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 Anterior Gradient Protein 2 (AGR2) 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) 80-99% 84-93% 94-105% 98-105%
EDTA plasma(n=5) 90-99% 92-104% 96-105% 80-103%
heparin plasma(n=5) 83-104% 80-89% 78-90% 94-101%

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 Anterior Gradient Protein 2 (AGR2)

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 Anterior Gradient Protein 2 (AGR2). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Anterior Gradient Protein 2 (AGR2). 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 Anterior Gradient Protein 2 (AGR2), 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 Anterior Gradient Protein 2 (AGR2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reference

  • Cancer BiomarkersSerum AGR2 as an early diagnostic and postoperative prognostic biomarker of human lung adenocarcinomaMetaPress: n7137r6271u54118
  • Molecular & Cellular ProteomicsIntegrated Proteomic Profiling of Cell Line Conditioned Media and Pancreatic Juice for the Identification of Pancreatic Cancer BiomarkersPubMed: PMC3205865
  • Prostate.Anterior gradient 2 (AGR2): blood-based biomarker elevated in metastatic prostate cancer associated with the neuroendocrine phenotypePubmed: 22911164
  • BMC Cancer.Validation of four candidate pancreatic cancer serological biomarkers that improve the performance of CA19.9Pubmed: 24007603
  • Clinical Cancer ResearchValidation of Biomarkers That Complement CA19.9 in Detecting Early Pancreatic CancerPubmed:25239611
  • PLoS OneAGR3 in Breast Cancer: Prognostic Impact and Suitable Serum-Based Biomarker for Early Cancer DetectionPubMed: 25875093