Complement System MBL (Lectin) ELISA

February 20, 2018

Background

The complement system plays an essential role in chronic, autoimmune and infectious disease. There are three pathways of complement activation, namely the classical, the alternative and the MBL pathway.

Impaired complement activity causes humans to become susceptible to repetitive fulminant or severe infections and may contribute to development of autoimmune disease. Inappropriate activation of complement contributes to chronic inflammation and tissue injury.

Technical Information

TheMBL Complement assay combines principles of the hemolytic assay for complement activation with the use of labeled antibodies specific for neoantigen produced as a result of complement activation. The amount of neoantigen generated is proportional to the functional activity of complement pathways.

In the Complement Screen MP kit the wells of the microtitre strips are coated with specific activators of the MBL pathway. Serum is diluted in diluent containing specific blocker to ensure that only the MBL pathway is activated. During the incubation of the subject's serum in the wells, complement is activated by the specific coating.

The wells are then washed and C5b-9 is detected with a specific alkaline phosphatase labelled antibody to the neoantigen expressed during MAC formation.

After a further washing step, detection of specific antibodies is obtained by incubation with alkaline phosphatase substrate solution. The amount of complement activation correlates with the colour intensity and is measured in terms of absorbance (optical density (OD))

Kit Components and Storage

- One frame with green break-apart wells (12x8) sealed in a foil pack with a desiccation sachet. The wells have been coated with mannan.

- 35 mL Diluent MP (Dil MP), labelled green.

- 13 mL conjugate containing alkaline phosphatase-labelled antibodies to C5b-9 (blue colour).

- 13 mL Substrate solution ready to use.

- 30 mL wash solution 30x concentrated.

- 0,2 mL negative control (NC) containing human serum. (to be diluted as for a subject's serum sample).

- 0,2 mL positive control (PC) containing freezed dried human serum, see “Reconstitution of positive control”, IFU

All reagents in the kit are ready for use except washing solution and controls. The reagents should be stored at 2-8° C except the positive control.

The positive control should be stored at -20° C.

Complement System MBL (Lectin) ELISA

Complement System MBL (Lectin) ELISA

February 20, 2018

Background

The complement system plays an essential role in chronic, autoimmune and infectious disease. There are three pathways of complement activation, namely the classical, the alternative and the MBL pathway.

Impaired complement activity causes humans to become susceptible to repetitive fulminant or severe infections and may contribute to development of autoimmune disease. Inappropriate activation of complement contributes to chronic inflammation and tissue injury.

Technical Information

TheMBL Complement assay combines principles of the hemolytic assay for complement activation with the use of labeled antibodies specific for neoantigen produced as a result of complement activation. The amount of neoantigen generated is proportional to the functional activity of complement pathways.

In the Complement Screen MP kit the wells of the microtitre strips are coated with specific activators of the MBL pathway. Serum is diluted in diluent containing specific blocker to ensure that only the MBL pathway is activated. During the incubation of the subject's serum in the wells, complement is activated by the specific coating.

The wells are then washed and C5b-9 is detected with a specific alkaline phosphatase labelled antibody to the neoantigen expressed during MAC formation.

After a further washing step, detection of specific antibodies is obtained by incubation with alkaline phosphatase substrate solution. The amount of complement activation correlates with the colour intensity and is measured in terms of absorbance (optical density (OD))

Kit Components and Storage

- One frame with green break-apart wells (12x8) sealed in a foil pack with a desiccation sachet. The wells have been coated with mannan.

- 35 mL Diluent MP (Dil MP), labelled green.

- 13 mL conjugate containing alkaline phosphatase-labelled antibodies to C5b-9 (blue colour).

- 13 mL Substrate solution ready to use.

- 30 mL wash solution 30x concentrated.

- 0,2 mL negative control (NC) containing human serum. (to be diluted as for a subject's serum sample).

- 0,2 mL positive control (PC) containing freezed dried human serum, see “Reconstitution of positive control”, IFU

All reagents in the kit are ready for use except washing solution and controls. The reagents should be stored at 2-8° C except the positive control.

The positive control should be stored at -20° C.

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