Q: We wish to measure MASP-2 levels in human plasma. Are stored plasma (or serum) at -20 centigrade stabile for this measurement if only frozen once. Should we measure in duplicate or triplicate? Is plasma better than serum as I have both stored for the MASP-2 test? What would be the cost of a kit and how many samples would it measure? I assume there is no special equipment other than ELISA like stuff? What is the shelf life as I have holiday next month? Is the a list of equipment we need to run the kits so I can check it with our lab technicians.

A: Once frozen samples should be fine for measurement. Generally, all experiments should be performed at least in triplicate so you can perform statistical analysis. MASP-2 should be detectable in both serum and plasma. Serum might yield cleaner signals since it is depleted of blood cells and clotting factors, but the depletion could, theoretically remove some MSAP-2 as well. Either type of sample should be suitable though. We currently have no MASP2 ELISA kits available. A list of our MASP2 antibodies can be found at this link if you would like to make your own. We guarantee all of our antibodies for 6 months from the receipt date.

MASP2 Overexpression Lysate

Novus Biologicals | Catalog # NBP2-05582

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for MASP2 Overexpression Lysate

MASP2 Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC218844

Accession#: NM_139208

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Theoretical MW

19 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for MASP2 Overexpression Lysate

Western Blot: MASP2 Overexpression Lysate [NBP2-05582]

Western Blot: MASP2 Overexpression Lysate (Adult Normal) [NBP2-05582] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for MASP2.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: MASP2

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]

Alternate Names

EC 3.4.21, EC 3.4.21.104, mannan-binding lectin serine peptidase 1 pseudogene 1, mannan-binding lectin serine peptidase 2, mannan-binding lectin serine protease 1 pseudogene 1, mannan-binding lectin serine protease 2, Mannose-binding protein-associated serine protease 2, MAP19, MASP1P1, MASP-2, MBL-associated plasma protein of 19 kD, MBL-associated serine protease 2, small MBL-associated protein, sMAP

Gene Symbol

MASP2

Additional MASP2 Products

Product Documents for MASP2 Overexpression Lysate

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Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for MASP2 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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