Human MASP2 (Mannan Associated Serine Protease 2) ELISA Kit (HUES03258)



ELISA Kit Technical ManualMSDS

Human MASP2 (Mannan Associated Serine Protease 2) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human MASP2 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human MASP2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human MASP2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human MASP2. The concentration of Human MASP2 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Assay time4.5h
Detection MethodColormetric
Detection Range3.13-200 ng/mL
Sensitivity1.88 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids


This kit recognizes Human MASP2 in samples. No significant cross-reactivity or interference between Human MASP2 and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

O.D Average Corrected


Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human MASP2 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human MASP2 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Mean (ng/mL)10.3117.8976.6010.0517.9883.08
Standard deviation0.600.733.070.581.064.25
C V (%)5.824.084.015.775.905.12


The recovery of Human MASP2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)92-10597
EDTA plasma (n=5)90-10497
Cell culture media (n=5)87-10094


Samples were spiked with high concentrations of Human MASP2 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)87-10187-10090-104
Average (%)929395
1:4Range (%)88-9981-9588-102
Average (%)938795
1:8Range (%)86-10180-9289-100
Average (%)938595
1:16Range (%)91-10985-10089-101
Average (%)999294

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human MASP2 (Mannan Associated Serine Protease 2) ELISA Kit (HUES03258) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Human MASP2 (Mannan Associated Serine Protease 2) ELISA Kit (HUES03258) Protein Information

UniProt Protein Function:MASP2: Serum protease that plays an important role in the activation of the complement system via mannose-binding lectin. After activation by auto-catalytic cleavage it cleaves C2 and C4, leading to their activation and to the formation of C3 convertase. Defects in MASP2 are the cause of MASP2 deficiency (MASPD). MASPD is a disorder that results in autoimmune manifestations, recurrent severe infections, and chronic inflammatory disease. Belongs to the peptidase S1 family. 2 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:EC; Protease; Secreted, signal peptide; Secreted

Chromosomal Location of Human Ortholog: 1p36.3-p36.2

Cellular Component: extracellular region

Molecular Function:peptidase activity; protein binding; serine-type endopeptidase activity; complement component C4b binding; calcium ion binding; calcium-dependent protein binding

Biological Process: innate immune response; complement activation, lectin pathway; proteolysis; complement activation, classical pathway; complement activation

Disease: Masp2 Deficiency

NCBI Summary:The Ra-reactive factor (RARF) is a complement-dependent bactericidal factor that binds to the Ra and R2 polysaccharides expressed by certain enterobacteria. Alternate splicing of this gene results in two transcript variants encoding two RARF components that are involved in the mannan-binding lectin pathway of complement activation. The longer isoform is cleaved into two chains which form a heterodimer linked by a disulfide bond. The encoded proteins are members of the trypsin family of peptidases. [provided by RefSeq, Jul 2008]
UniProt Code:O00187
NCBI GenInfo Identifier:21264363
NCBI Gene ID:10747
NCBI Accession:NP_006601.2
UniProt Secondary Accession:O00187,O75754, Q5TEQ5, Q5TER0, Q96QG4, Q9BZH0, Q9H498 Q9H499, Q9UBP3, Q9UC48, A8K458, A8MWJ2,
UniProt Related Accession:O00187
Molecular Weight:76 kDa
NCBI Full Name:mannan-binding lectin serine protease 2 isoform 1 preproprotein
NCBI Synonym Full Names:mannan-binding lectin serine peptidase 2
NCBI Official Symbol:MASP2
NCBI Official Synonym Symbols:sMAP; MAP19; MASP-2; MASP1P1
NCBI Protein Information:mannan-binding lectin serine protease 2
UniProt Protein Name:Mannan-binding lectin serine protease 2
UniProt Synonym Protein Names:MBL-associated serine protease 2; Mannose-binding protein-associated serine protease 2; MASP-2
Protein Family:Mannan-binding lectin serine protease
UniProt Gene Name:MASP2
UniProt Entry Name:MASP2_HUMAN
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