Multiplex Assay Kit for Anterior Gradient 2 (AGR2) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Anterior Gradient 2 (AGR2) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Anterior Gradient 2 (AGR2) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 88-96 92
EDTA plasma(n=5) 79-91 84
heparin plasma(n=5) 91-102 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anterior Gradient 2 (AGR2) ,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 Anterior Gradient 2 (AGR2) ,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 Anterior Gradient 2 (AGR2) ,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) 80-95% 88-95% 89-96% 78-97%
EDTA plasma(n=5) 84-98% 78-95% 83-101% 78-94%
heparin plasma(n=5) 88-95% 87-96% 90-103% 79-88%

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#:AGR2) 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
Cancer Biomarkers Serum AGR2 as an early diagnostic and postoperative prognostic biomarker of human lung adenocarcinoma MetaPress: n7137r6271u54118
Molecular & Cellular Proteomics Integrated Proteomic Profiling of Cell Line Conditioned Media and Pancreatic Juice for the Identification of Pancreatic Cancer Biomarkers PubMed: PMC3205865
Prostate. Anterior gradient 2 (AGR2): blood-based biomarker elevated in metastatic prostate cancer associated with the neuroendocrine phenotype Pubmed: 22911164
BMC Cancer. Validation of four candidate pancreatic cancer serological biomarkers that improve the performance of CA19.9 Pubmed: 24007603
Clinical Cancer Research Validation of Biomarkers That Complement CA19.9 in Detecting Early Pancreatic Cancer Pubmed:25239611
PLoS One AGR3 in Breast Cancer: Prognostic Impact and Suitable Serum-Based Biomarker for Early Cancer Detection PubMed: 25875093
Clinical Neurology and Neurosurgery Serum AGR2 as a useful biomarker for pituitary adenomas S0303846717300045
Oncotarget Identification, characterization and application of a new peptide against anterior gradient homolog 2 (AGR2) Pubmed:29937991
Int J Oncol Anterior gradient protein 2 is a marker of tumor aggressiveness in breast cancer and favors chemotherapy‑induced senescence escape 34913074
International Journal of Cancer Proteome profiling of enzalutamide‐resistant cell lines and serum analysis identified ALCAM as marker of resistance in castration‐resistant prostate cancer Pubmed:35689436
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
RPC285Hu01 Recombinant Anterior Gradient 2 (AGR2) Positive Control; Immunogen; SDS-PAGE; WB.
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