Multiplex Assay Kit for Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

DGSX; OCI5; SDYS; SGB; SGBS1; MXR7; Glypican Proteoglycan 3; GTR2-2; Intestinal protein OCI-5; Secreted glypican-3

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

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  • Multiplex Assay Kit for Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA971Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-101 91
EDTA plasma(n=5) 95-102 98
heparin plasma(n=5) 92-99 96
sodium citrate plasma(n=5) 79-93 82

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glypican 3 (GPC3) ,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 Glypican 3 (GPC3) ,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 Glypican 3 (GPC3) ,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) 98-105% 96-105% 78-101% 94-101%
EDTA plasma(n=5) 94-105% 83-103% 87-99% 85-104%
heparin plasma(n=5) 93-102% 90-105% 89-96% 85-104%
sodium citrate plasma(n=5) 84-93% 82-89% 83-97% 78-92%

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#:GPC3) 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.

Multiplex Assay Kit for Glypican 3 (GPC3) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Diagnostic Role of Serum Glypican-3 as a Tumor Marker for Hepatocellular Carcinoma Sciencepub:10043238
  • Cytotoxic effects of antiglypican-3 against HepG2 cell lineEbscohost: Source
  • Can Glypican3 be Diagnostic for Early Hepatocellular Carcinoma among Egyptian Patients?Pubmed: 24460300
  • Diagnostic accuracy of serum glypican-3 for hepatocellular carcinoma: A systematic review and meta-analysisPubmed: 24362268
  • Diagnostic Role of Serum Glypican-3 as Hepatocellular CarcinomaZolcat: Source
  • Role of serum glypican-3 in the diagnosis and differentiation of small hepatocellular carcinoma from hepatitis-C virus cirrhosisScienceDirect: S2090506814000062
  • Comparing prothrombin induced by vitamin K absence-II (PIVKA-II) with the oncofetal proteins glypican-3, Alpha feto protein and carcinoembryonic antigen in diagnosing hepatocellular carcinoma among Egyptian patients.Pubmed:24841158
  • Evaluation of antiglypican-3 therapy as a promising target for amelioration of hepatic tissue damage in hepatocellular carcinomaPubmed:25449037
  • Predictive value of tumor markers in patients with recurrent hepatocellular carcinoma in different vascular invasion patternabstract:abstract4525.shtml
  • Glypican-3 level assessed by the enzyme-linked immunosorbent assay is inferior to alpha-fetoprotein level for hepatocellular carcinoma diagnosispmc:articles
  • Glypican‐3: A promising biomarker for hepatocellular carcinoma diagnosis and treatmentpubmed:28621802
  • Cytotoxic and partial hepatoprotective activity of sodium ascorbate against hepatocellular carcinoma through inhibition of sulfatase-2 andPubmed:29669302
  • Development and Validation of a Two-Site Immunoradiometric assay for Glypican-3 in plasma: Implication in Diagnosis of Hepatocellular CarcinomaResearchgate:Source
  • Serum Glypican-3, Vascular Endothelial Growth Factor, and Interleukin-6 Levels in Hepatocellular Carcinoma10.5505:tjo.2018.1742
  • Elevated plasma glypicans are associated with organ failure in patients with infectionPubmed: 30618011
  • VALUE OF SERUM GLYPICAN 3 AS A PREDICTIVE TOOL FOR EARLY DIAGNOSIS OF HEPATOCELLULAR CARCINOMA (HCC)
  • Search for Effective Serum Tumor Markers for Early Diagnosis of Hepatocellular Carcinoma Associated with Hepatitis C
  • Selenium nanoparticles overcomes sorafenib resistance in thioacetamide induced hepatocellular carcinoma in rats by modulation of mTOR, NF-κB pathways and …Pubmed:35640776

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