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PNS14 DIVER and PNS14 Line Assay

PNS14 DIVER and PNS14 Line Assay
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MAG DIVER and Gangliosid Line Assay
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ravo Stiff-Person Blot

Recombinant immunoblot for the detection of GAD65- and Amphiphysin-autoantibodies for the diagnosis of
Stiff-Person-Syndrome


Stiff-Person Syndrome is a rare neurological autoimmune disease that is character-ized by rigidity and episodic spasms of muscles as a result of continuous motor unit activity.
Antibodies to GAD (glutamat-decarboxylase) are considered as serological markers for Stiff-Person-Syndrome. The majority of patients have high titers of antibodies to both isoforms, GAD65 and GAD67. These enzymes catalyse the conversation of glutamate to GABA (ã-aminobutyric acid), a major inhibitory neurotransmitter of the CNS.
Amphiphysin expression is found in synaptic vesicels of neurons as well as in the sceletal musculature. The presence of anti-Amphiphysin antibodies (with or without GAD) indicates a paraneoplastic neurological syndrome caused by an underlying tumour.


Procedure (Short Description):

• Cover one strip with the ready to use positive and negative control (2 ml).

• Cover the other strips with 1.5 ml ready to use dilution buffer.

• Add 15 µl of specimen and mix  carefully (end-dilution: 1 : 100).

• Incubate for 60 minutes at room temperature on a rocking table.

• Wash five times with diluted wash buffer.

• Add the ready to use conjugate (2ml) for the positive control to the respective
  strip. Specimen and negative control: add 2 ml alkaline phosphatase IgG
  conjugate, ready to use, per strip.

• Incubate for 30 minutes at room temperature on a rocking table.

• Wash five times with diluted wash buffer.

• Cover each strip with 2 ml ready to use substrate-solution.

• Incubate for 25 minutes at room temperature until the bands become clearly
   visible. See control scan for comparison.

• Transfer the strips to distilled water to stop the reaction. Put the strips onto
   filter paper and let them dry. Store the strips in the dark.

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ravo Diagnostika, Oltmannsstr. 5, D-79100 Freiburg
phone +49 761 40 74 88, fax +49 761 40 74 77, email