ELISA Kit for Gelsolin (GSN)

GS; AGEL; ADF; Brevin; Gelsolin Amyloidosis,Finnish Type; Actin-depolymerizing factor

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 85-94 91
EDTA plasma(n=5) 95-103 99
heparin plasma(n=5) 78-99 80

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Gelsolin (GSN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Gelsolin (GSN) 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 Gelsolin (GSN) 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) 93-101% 84-98% 80-101% 88-96%
EDTA plasma(n=5) 81-90% 78-94% 96-103% 87-101%
heparin plasma(n=5) 78-98% 81-102% 85-101% 96-105%

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
Neurochmical Research Gelsolin in Cerebrospinal Fluid as a Potential Biomarker of Epilepsy SpringerLink: 3585r8h46q7r0171
Journal of Neuroinflammation Treatment with gelsolin reduces brain inflammation and apoptotic signaling in mice following thermal injury BioMed: 17422094
Biomedical Engineering and Informatics Application of iTRAQ quantitative proteomics in identification of serum biomarkers in breast cancer IEEE: 6098563
European Journal of Obstetrics & Gynecology and Reproductive Biology Investigation of the actin scavenging system in pre-eclampsia Pubmed: 24239294
The Annals of Thoracic Surgery Proteomic Study Reveals Plasma Protein Changes in Congenital Heart Diseases Pubmed: 24565402
BMC Anesthesiol.? The role of plasma gelsolin in cardiopulmonary bypass induced acute lung injury in infants and young children: a pilot study Pubmed:Pmc4132929
Cell Biochemistry and Biophysics Gelsolin: Role of a Functional Protein in Mitigating Radiation Injury Pubmed:25164111
Neurosci Lett Plasma gelsolin and matrix metalloproteinase 3 as potential biomarkers for Alzheimer disease PubMed: 25864780
Proteomics Clinical Applications Comparative mass spectrometric and immunoassay‐based proteome analysis in serum of Duchenne muscular dystrophy patients Pubmed:26680509
PROTEOMICS - Clinical Applications Comparative mass spectrometric and immunoassay-based proteome analysis in serum ofDuchenne muscular dystrophy patients. pubmed:26680509
Medicina Intensiva Serum gelsolin levels in aneurismal subarachnoid hemorrhage: Preliminary results. pubmed:28365030
Journal of Alzheimers Disease Urine-Based Biomarkers for Alzheimer's Disease Identified Through Coupling Computational and Experimental Methods Pubmed:30040720
World Journal of Pediatrics Plasma gelsolin level predicts acute kidney injury after cardiopulmonary bypass in infants and young children Pubmed:29427164
Biomedicines Production and Secretion of Gelsolin by Both Human Macrophage-and Fibroblast-like Synoviocytes and GSN Modulation in the Synovial Fluid of Patients with Various … Pubmed:35327525
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