Description: MASP-3, Human, ELISA kit

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.

Specifications:

Catalog number Product type Quantity Standard range Detection level Working volume Species Application Disease Principle Linearity Recovery Storage and stability Precautions References

HK339-01

Assays

1 x 96 det.

8 – 500 ng/ml

8 ng/ml

100 µl/well

Human

The human MASP-3 ELISA kit is to be used for the in vitro quantitative determination of both natural and recombinant human MASP-3 in serum and plasma samples.

Autoimmunity, Infectious diseases

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.

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.

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%).

Product should be stored at 4 °C. Under recommended storage conditions, product is stable for one year.

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.

1. Frauenknecht, V et al; Plasma levels of mannan-binding lectin (MBL)-associated serine proteases (MASPs) and MBL-associated protein in cardio- and cerebrovascular diseases. Clinical & Experimental Immunology 2013

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