ELISA Kit for Serglycin (SRGN)

PRG; PPG; PRG1; proteoglycan 1, secretory granule

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 83-96 88
EDTA plasma(n=5) 78-98 89
heparin plasma(n=5) 83-104 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Serglycin (SRGN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Serglycin (SRGN) 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 Serglycin (SRGN) 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) 88-102% 91-98% 90-97% 85-101%
EDTA plasma(n=5) 80-93% 82-99% 85-92% 90-104%
heparin plasma(n=5) 92-99% 89-96% 87-101% 79-99%

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.

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Magazine Citations
Irish Journal of Medical Science The role of increased synovial fluid A disintegrin and metalloproteinase with thrombospondin motifs4 and serglycin levels in osteoarthritis Pubmed: 30536194
MOLECULAR AND CELLULAR BIOCHEMISTRY SRGN, a new identified shear-stress-responsive gene in endothelial cells Pubmed: 32712749
Theranostics Significance of serglycin and its binding partners in autocrine promotion of metastasis in esophageal cancer 33456569
CELLULAR ONCOLOGY Cancer©\associated fibroblasts secrete hypoxia©\induced serglycin to promote head and neck squamous cell carcinoma tumor cell growth in vitro and in vivo by ¡­ 33651283
Cell Death Dis Serglycin induces osteoclastogenesis and promotes tumor growth in giant cell tumor of bone 34556636
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