MASP-3, Human, ELISA kit - HK339

Innate Immunity

Inflammation

Cell- and tissue damage

Name

MASP-3, Human, ELISA kit

Catalog nr

HK339 (lot number and expiry date are indicated on the label)

Short description

Size

2 x 96 det.

Application

Manual

HK339.pdf

Description

The mannan-binding lectin (MBL) pathway of the complement system is initiated by the binding of MBL or ficolin to specific carbohydrate structures found on the surface of a variety of microorganisms. This binding is followed by the activation of MBL-associated serine proteases (MASPs). There are four known MASPs: MASP-1, MASP-2, MASP-3 and Map19 (also known as sMAP, a small, separately synthesized fragment of MASP-2). MASPs are secreted as single chain precursors and are processed into a heavy chain (A) and catalytic light chain (B) linked by disulfide bonds. All MASPs form homo-dimers and individually associate with MBL and the ficolins in a Ca2+-dependent manner.

MASP-3 is an alternatively spliced product from the MASP-1 gene, containing an identical A chain, but an entirely different B chain. MASP-3 is an approximately 100 kDa protein and associates with both MBL and L-ficolin in the presence of Ca2+. The MASP3 dimer can be disrupted by EDTA.

MASP-3 is expressed mainly but not exclusively in the liver. Liver expressed MASP-3 is secreted into the blood. Unlike MASP-2, no proteolytic activity on complement proteinss C2, C3 and C4 for MASP-3 has been detected. MASP-3 exerts inhibitory activity on MASP-2 function. However, no natural substrates for MASP-3 are known.

MASP-3 concentration in healthy human serum is normally distributed with a mean value of 3796 ng/ml. MASP-3 is present at birth at approximately 80% of the level measured 6, 9 and 12 months after birth. The MASP-3 level is stable throughout a 1-year period.

The high expression of MASP-3 in astrocytes indicates that MASP-3 might also be involved in other functions that could be important in the brain. In multiple sclerosis patients, high MASP-3 levels seem to have a positive effect (median MASP-3 levels: 4539 ng/ml). MASP-3 deficiencies are currently not reported.

Cross Reactivity

Potential cross reacting proteins detected in the MASP-3 ELISA.

Cross reactant

Reactivity

MASP-1

MASP-2

Negative

Application

The human MASP-3 ELISA kit is to be used for the in vitro quantitative determination of both natural and recombinant human MASP-3. This kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures. In serum or plasma samples MASP-3 can be measured accurately if serum or plasma samples are diluted at least 20 times. Most reliable results are obtained if EDTA plasma is used.

Typical standard curve

Principle

The MASP-3 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 4½ hours.
The efficient format of 2 plates with twelve disposable 8-well strips allows free choice of batch size for the assay.
Samples and standards are captured by a solid bound specific antibody.
Biotinylated tracer antibody will bind to captured MASP-3.
Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody.
Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB).
The enzyme reaction is stopped by the addition of oxalic acid.
The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the MASP-3 standards (log).
The MASP-3 concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Storage and stability

Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least six months. After reconstitution the reagents are stable for 1 month if stored at 2-8°C.

Recovery

Normal human blood samples, containing baseline levels of MASP-3, were spiked with recombinant MASP-3 in concentrations of 100 and 10 ng/ml. Samples with and without MASP-3 were incubated for 30 minutes at 37°C. Samples were measured using the ELISA. Values for MASP-3 ranged between 93% and 116% (mean 110%).

Linearity

The linearity of the assay was determined by serially diluting a sample containing 250 ng/ml human MASP-3. The diluted samples were measured in the assay. The line obtained a slope of 0.970 and a correlation coefficient of 0.999.

Precautions

For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.

Also available

Scientific info

ELISA to elucidate the role of human MASP-3
The mannan-binding lectin (MBL) pathway of the complement system is initiated by the binding of MBL or ficolin to specific carbohydrate structures found on the surface of a variety of microorganisms. This binding is followed by the activation of MBL-associated serine proteases (MASPs).
MASP-3 is an alternatively spliced product from the MASP-1 gene, containing an identical A chain, but an entirely different B chain. MASP-3 is an approximately 100 kD protein and associates with both MBL and L-ficolin in the presence of Ca2+. The MASP-3 dimer can be disrupted by EDTA.

MASP-3 function
Unlike MASP-2, no proteolytic activity on complement proteins C2, C3 and C4 for MASP-3 has been detected. MASP-3 exerts inhibitory activity on MASP-2 function. However, no natural substrates of MASP-3 are known.

MASP-3 expression
MASP-3 is expressed mainly but not exclusively in the liver and is secreted into the blood. The high expression of MASP-3 in astrocytes indicates that MASP-3 might also be involved in other functions that could be important in the brain.
MASP-3 concentration in healthy human blood is normally distributed with a mean value of 3796 ng/ml. MASP-3 is present at birth at approximately 80% of the level measured 6, 9 and 12 months after birth. The MASP-3 level is stable throughout a 1-year period. The role of MASP-3 in biological processes is not clear. In multiple sclerosis (MS) patients, MASP-3 levels seem to be negatively correlated with MS-associated human endogenous retrovirus HERV-H (median MASP-3 levels: 4539 ng/ml). MASP-3 deficiencies have not yet been reported.

Special features of the assay
• Tool to elucidate the biological role of MASP-3
• Unique assay
• Reliable and fast technique
• Measurable concentration 8 - 500 ng/ml
• Detection limit 8 ng/ml

1. Thiel, S. et al; An assay for mannan-binding lectin (MBL) associated serine protease 3, MASP-3
2. Zundel, S. et al; Characterization of recombinant mannan-binding lectin-associated serine protease (MASP)-3 suggests an activation mechanisms different from that of MASP-1 and MASP-2. J Immunol 2004, 172: 4342
3. Moller-Kristensen, M. et al; Cooperation between MASP-1 and MASP-2 in the generation of C3 convertase through the MBL pathway. Int Immunol 2006, 19: 141
4. Kuraya, M. et al; Expression of H-ficolin/Hakata antigen, mannose-binding lectin-associated serine protease (MASP)-1 and MASP-3 by human glioma cell line T98G. Int Immunol 2003, 15: 109
5. Sorensen, R. et al; Mannan-binding-lectin-associated serine protease, characteristics and disease associations. Springer Semin Immun 2005, 27: 299
6. Christensen, T. et al; Gene-environment interactions in multiple sclerosis: innate and adaptive immune responses to human endogenous retrovirus and herpesvirus antigens and the lectin complement activation pathway. J Neuroimmunol 2007, 183: 175

MSDS

MSDS ELISA kit.pdf

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