ELISA Kit for Mannose Associated Serine Protease 1 (MASP1)

CRARF; PRSS5; Mannan-Binding Lectin Serine Peptidase 1; Ra-reactive factor serine protease p100; Complement-activating component of Ra-reactive factor; Serine protease 5

Specificity

This assay has high sensitivity and excellent specificity for detection of Mannose Associated Serine Protease 1 (MASP1).
No significant cross-reactivity or interference between Mannose Associated Serine Protease 1 (MASP1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mannose Associated Serine Protease 1 (MASP1) and the recovery rates were calculated by comparing the measured value to the expected amount of Mannose Associated Serine Protease 1 (MASP1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-99 85
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 84-98 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mannose Associated Serine Protease 1 (MASP1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mannose Associated Serine Protease 1 (MASP1) 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 Mannose Associated Serine Protease 1 (MASP1) 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) 94-103% 91-105% 91-99% 83-94%
EDTA plasma(n=5) 89-99% 84-104% 84-97% 78-97%
heparin plasma(n=5) 91-98% 88-95% 80-90% 96-103%

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
Pathology Increased Circulating Plasma Mannose Binding Lectin Associated Serine Protease (Masp) As A Result of Enu-Induced Mutation in Masp-1 Lww: Source
The Journal fo Biological Chemistry Bisretinoid-mediated Complement Activation on Retinal Pigment Epithelial Cells Is Dependent on Complement Factor H Haplotype Pubmed:24550392
the journal of immunology The Levels of the Lectin Pathway Serine Protease MASP-1 and Its Complex Formation with C1 Inhibitor Are Linked to the Severity of Hereditary Angioedema PubMed: 26371246
J Neuroinflammation.  Mannose-binding lectin-associated serine protease 2 (MASP-2) contributes to poor disease outcome in humans and mice with pneumococcal meningitis PMC5234106
Frontiers in Immunology AmpliSeq screening of genes encoding the C-type lectin receptors and their signaling components reveals a common variant in MASP1 associated with … Pubmed:29515573
Journal of Proteomics Plasma proteome profiling of high-altitude polycythemia using TMT-based quantitative proteomics approach Pubmed: 30605725
Kidney International Reports Clinical value of complement activation biomarkers in overt diabetic nephropathy
Kidney International Reports Comparison of Complement Pathway Activation in Autoimmune Glomerulonephritis Pubmed:35571000
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