Membrane Cofactor Protein ELISA Kit (MCP/CD46)

INTENDED USE

Human MCP/CD46 ELISA kit can measure concentrations of MCP/CD46 (Membrane Cofactor Protein, MCP, CD46, complement regulatory protein, trophoblast leucocyte common antigen, AHUS2, MIC10) present is plasma, serum, biological fluids or tissue homogenate samples. This assay has a minimum sensitivity detection limit of 4.688 pg/ml.

INTRODUCTION

Membrane cofactor protein (MCP) also referred to as CD46 plays a role, in regulating the activation of complement to safeguard host cells from complement related damage. It is a transmembrane glycoprotein that can be found widely throughout the body and has the ability to bind with complement components C3b and C4b. By acting as a cofactor it aids in their breakdown by serum factor I.

Through its promotion of the inactivation of C3b and C4b, MCP prevents these complement opsonins from coming to form the C3 and C5 convertases. As a result it effectively hampers the progression of the complement cascade. This restriction ensures that complement activation occurs on cell surfaces shielding host cells from harm caused by complement mediated attacks. MCPs significance is particularly evident on host cells that lack complement regulators like erythrocytes, platelets and fetal trophoblasts. Additionally it provides protection against the deposition of complements triggered by autoreactive antibodies in diseases. By virtue of its functions within the complement system it acts as a marker that distinguishes between self (host) and foreign cells.

In addition to its role in limiting complement activity it also participates in signaling processes that influence cell behavior. When pathogens or inflammatory molecules bind with MCP signals are triggered which impact dendritic cell maturation T cell polarization and macrophage phagocytosis. Mice lacking MCPs demonstrate deficiencies, in cell function and CD4+ T cell immunity.

Given its role, in regulating the system there is a growing interest in using MCP as a target for therapeutic purposes. In the laboratory animal studies scientists have found that it can effectively prevent the activation of complement, which is crucial in autoimmune diseases and organ transplantation. On the one hand by blocking the signals of MCP it may be possible to reduce inflammation by influencing how dendritic cells and macrophages respond. Moreover since many harmful microorganisms rely on MCPs to invade host cells researchers are exploring ways to disrupt this interaction and prevent infections.

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- MCP/CD46.
• 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 MCP/CD46 ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 7.812, 15.625, 31.25, 62.5, 125, 250, 500 pg/ml.
Reactivity: Human
Sensitivity: 4.688 pg/ml
Range: 7.813 – 500 pg/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for MCP/CD46, no cross reactivity or interference between MCP/CD46 and analogues was detected.
– Recovery: Serum (93 – 99%), EDTA Plasma (86 – 105%), Heparin Plasma (90 – 103%).
– Linearity: Serum (86 – 99%), EDTA Plasma (87 – 98%), Heparin Plasma (88 – 97%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. Protective effect of membrane cofactor protein against complement-dependent injury. Acta Pharmacol Sin. (2005) 26 (8): 987-91. Xu D., et al.
2. CD46: the ‘multitasker’ of complement proteins. Int J Biochem Cell Biol. (2013) 45 (12): 2808-20. Yamamoto H., et al.
3. Emerging roles and new functions of CD46. Springer Semin Immunopathol. (2005) 27 (3): 345-58. Liszewski M.K., et al.
4. The molecular biology of thrombotic microangiopathy. Kidney Int. (2006) 70 (1): 16-23. Tsai H.M.

ADDITIONAL INFORMATION

• Full Name: Membrane Cofactor Protein ELISA Kit (MCP/CD46)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 4.688 pg/ml

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