Multiplex Assay Kit for Golgi Phosphoprotein 2 (GOLPH2) ,etc. by FLIA (Flow Luminescence Immunoassay)

GOLM1; GP73; C9orf155; Golgi Membrane Protein 1; Golgi Protein 73

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Golgi Phosphoprotein 2 (GOLPH2) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Golgi Phosphoprotein 2 (GOLPH2) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Golgi Phosphoprotein 2 (GOLPH2) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Golgi Phosphoprotein 2 (GOLPH2) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-94 89
EDTA plasma(n=5) 83-90 86
heparin plasma(n=5) 83-102 98
sodium citrate plasma(n=5) 78-104 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Golgi Phosphoprotein 2 (GOLPH2) ,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 Golgi Phosphoprotein 2 (GOLPH2) ,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 Golgi Phosphoprotein 2 (GOLPH2) ,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) 78-103% 82-101% 84-103% 82-90%
EDTA plasma(n=5) 89-96% 96-105% 93-105% 85-104%
heparin plasma(n=5) 90-98% 94-102% 86-96% 83-91%
sodium citrate plasma(n=5) 90-104% 87-98% 85-99% 79-96%

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#:GOLPH2) 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
Molecular & Cellular Proteomics LAMC2: A promising new pancreatic cancer biomarker identified by proteomic analysis of pancreatic adenocarcinoma tissues Mcponline: 023507
Afro-Egypt J Infect Endem Dis Diagnostic and Prognostic Validity of Serum Golgi Protein 73 in Egyptian Patients with Hepatocelluar Carcinoma Ajied: Ajied_System_Files
Journal of Applied Pharmaceutical Science Diagnostic role of Golgi protein 73 and IL-17 in Egyptian cirrhotic rather than hepatocellular carcinoma patients. handle:123456789
Oncology Letters Serum Golgi protein 73 is a prognostic rather than diagnostic marker in hepatocellular carcinoma pubmed:29113278
Scientific Reports The possible role of Dickkopf-1, Golgi protein-73 and Midkine as predictors of hepatocarcinogenesis: a review and an Egyptian study Pubmed: 32198440
Hepatobiliary & Pancreatic Diseases International Dynamic expression of hepatic GP73 mRNA and protein and circulating GP73 during hepatocytes malignant transformation Pubmed: 32171652
Biomolecules Deoxycholic Acid Upregulates Serum Golgi Protein 73 through Activating NF-¦ÊB Pathway and Destroying Golgi Structure in Liver Disease. Biomolecules 2021, 11?¡­ 33540642
ADVANCED RESEARCHES Search for Effective Serum Tumor Markers for Early Diagnosis of Hepatocellular Carcinoma Associated with Hepatitis C
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