ELISA Kit for Mannose Associated Serine Protease 1 (MASP1) Mus musculus (Mouse) Sandwich ELISA

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

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  • ELISA Kit for Mannose Associated Serine Protease 1 (MASP1) Packages (Simulation)
  • ELISA Kit for Mannose Associated Serine Protease 1 (MASP1) Packages (Simulation)
  • ELISA Kit for Mannose Associated Serine Protease 1 (MASP1) Results demonstration
  • SEB895Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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.

ELISA Kit for Mannose Associated Serine Protease 1 (MASP1)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mannose Associated Serine Protease 1 (MASP1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mannose Associated Serine Protease 1 (MASP1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mannose Associated Serine Protease 1 (MASP1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Mannose Associated Serine Protease 1 (MASP1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Increased Circulating Plasma Mannose Binding Lectin Associated Serine Protease (Masp) As A Result of Enu-Induced Mutation in Masp-1Lww: Source
  • Bisretinoid-mediated Complement Activation on Retinal Pigment Epithelial Cells Is Dependent on Complement Factor H HaplotypePubmed:24550392
  • The Levels of the Lectin Pathway Serine Protease MASP-1 and Its Complex Formation with C1 Inhibitor Are Linked to the Severity of Hereditary AngioedemaPubMed: 26371246
  • Mannose-binding lectin-associated serine protease 2 (MASP-2) contributes to poor disease outcome in humans and mice with pneumococcal meningitisPMC5234106
  • AmpliSeq screening of genes encoding the C-type lectin receptors and their signaling components reveals a common variant in MASP1 associated with …Pubmed:29515573
  • Plasma proteome profiling of high-altitude polycythemia using TMT-based quantitative proteomics approachPubmed: 30605725
  • Clinical value of complement activation biomarkers in overt diabetic nephropathy
  • Comparison of Complement Pathway Activation in Autoimmune GlomerulonephritisPubmed:35571000

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