MASP2 ELISA Kit (Mannan-Binding Lectin Serine Protease 2)

INTENDED USE

Human MASP2 ELISA kit can measure concentrations of mannan-binding lectin serine protease 2 (MASP2, MBL-associated serine protease 2MASP-2) present is plasma, serum, biological fluids or tissue homogenate samples. This assay has a minimum sensitivity detection limit of 1.875 ng/ml.

INTRODUCTION

MASP2, known as mannan-binding lectin serine protease 2, is an enzyme playing a key role in the lectin pathway of the complement system within innate immunity. As a central component of the multiprotein MBL complex, it cleaves complement proteins to activate inflammatory and pathogen-targeting responses upon recognizing foreign carbohydrate surfaces. Dysregulation of its activity however can trigger autoimmune complications.

Produced in the liver then circulating in inactive form until binding MBL and ficolins, it becomes activated when its regulatory complex identifies pathogenic cell surface carbohydrates through pattern recognition. This prompts a conformational change in MASP2, allowing it to cleave complement proteins C4 and C2 into active fragments that trigger the downstream complement response cascade through their own proteolytic signaling on immune cell surfaces and pathogens. C4 cleavage additionally sets off the non-specific alternative complement pathway for amplifying the immune response.

By spurring this proteolytic cascade, MASP2 serves as the initiator and rate-limiting catalyst driving lectin pathway activation based on glycoprotein detection. The enzyme’s cleavage specificity and requirement for molecular configuration change upon substrate binding allows tight control over complement response activation to avoid unnecessary inflammation. However, dysfunctional regulation of MASP2 activity can lead to consumption of complement components even without pathogen triggers. This propagates inflammation contributing to autoimmune and ischemic injury diseases like rheumatoid arthritis and stroke.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti- MASP2
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this MASP2 ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 3.125, 6.25, 12.5, 25, 50, 100, 200 ng/ml.
Reactivity: Human
Sensitivity: 1.875 ng/ml
Range: 3.125 – 200 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for MASP2, no cross reactivity or interference between MASP2 and analogues was detected.
– Recovery: Serum (86 – 105%), EDTA Plasma (85 – 98%), Heparin Plasma (91 – 103%).
– Linearity: Serum (85 – 101%), EDTA Plasma (85 – 101%), Heparin Plasma (82 – 100%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. Mannan-binding-lectin-associated serine proteases, characteristics and disease associations. Springer Semin Immunopathol. (2005) 27 (3): 299-319. Sørensen R., et al.
2. MASP2 inhibition by narsoplimab suppresses endotheliopathies characteristic of transplant-associated thrombotic microangiopathy: in vitro and ex vivo evidence. Clin Exp Immunol. (2023) 213 (2): 252-264. Elhadad S., et al.
3. Narsoplimab for severe transplant-associated thrombotic microangiopathy. Thromb J. (2023) 21 (1): 26. Pandrowala A., et al.
4. Mannan-binding lectin and mannan-binding lectin-associated serine protease 2 in acute pancreatitis. Pancreas. (2011) 40 (7): 1097-102. Novovic S., et al.

ADDITIONAL INFORMATION

• Full Name: MASP2 ELISA Kit (Mannan-Binding Lectin Serine Protease 2)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 1.875 ng/ml

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