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CLIA Kit for Granulin (GRN)

GEP; GP88; PCDGF; PEPI; PGRN; Proepithelin; Acrogranin; Glycoprotein of 88 Kda; Paragranulin

Specificity

This assay has high sensitivity and excellent specificity for detection of Granulin (GRN).
No significant cross-reactivity or interference between Granulin (GRN) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Granulin (GRN) and the recovery rates were calculated by comparing the measured value to the expected amount of Granulin (GRN) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-102 98
EDTA plasma(n=5) 91-99 94
heparin plasma(n=5) 93-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Granulin (GRN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Granulin (GRN) 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 Granulin (GRN) 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) 99-105% 91-98% 80-105% 82-99%
EDTA plasma(n=5) 79-93% 80-97% 91-98% 87-105%
heparin plasma(n=5) 85-104% 96-103% 82-102% 90-104%

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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Endokrynologia Polska Serum progranulin levels are elevated in infertile women with obesity Pubmed: 30229554
Endokrynologia Polska Effects of serum granulin levels on anthropometric measures and glucose metabolism in infertile women with different ovarian reserve status Pubmed: 30845343
Sci Rep AZP2006, a new promising treatment for Alzheimer's and related diseases 34413330
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