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Oligoclonal bands in multiple sclerosis Patterns 1 5, described in this section, are typically common (but real examples).
The important patterns in MS are those that demonstrate "intrathecal IgG synthesis" (pattern 2 and 3). For the detection of oligoclonal bands, paired serum and CSF samples are required. Blood must be taken at the same time as the lumbar puncture (LP), if not then within two to three weeks of the LP. Results cannot be relied upon beyond this time limit (half life of serum IgG is about 23 days). This example demonstrates the effect of unpaired samples on oligoclonal bands. Serum was taken six weeks after the LP and contains extra bands which are not reflected in the CSF. The absence of bands 1970s louis vuitton purses in the serum corresponding to those present in the CSF might lead to incorrect interpretation of intrathecal IgG synthesis (pattern 2 and/or 3). Due to the time delay the relvant IgG may have been cleared from the serum. The principle involves the separation of proteins louis vuitton bags women&s handbags (IgG's) in the paired serum and cerebrospinal fluid (CSF) using agarose gel electrophoresis followed by passive transfer louis vuitton large ring agenda cover onto nitrocellulose membrane. CASTING GEL: AT 65 add 2ml (pH 3 10) 0.5ml (pH 8 10.5) pharmalyte to molten agarose (65 70 mix and cast at 65 HYSTERESIS: Keep the casted gel at 4 in a moist chamber for at least 30 min. SAMPLE PREPARATION: Serum diluted at 1:400 5 loaded. Vol. ( of CSF loaded = (2.5/total protein). RUNNING THE GEL: louis vuitton bags kuwait prices Blot the gel with nitrocellulose membrane (NCM). Position the electrode wicks at 7cm apart (1M NaOH ( ve), 0.05M H2SO4 (+ve)) on the gel and then the application strip 2cm from +ve electrode. Remove application strip after 20mins. The gel is electrophoresed at 1250 volt/hour. BLOTTING: Pre blot gel for 10 sec with NCM and discard it. Blot with another NCM piece for 30min. BLOCKING: Dry the blot.
Incubate in 2% marvel/saline for 30minutes and quick rinse in tap water. PRIMARY ANTIBODY: Anti human IgG (gamma chain) peroxidase, 1:200 dilution in 0.2% marvel/saline and incubate for 60mins.
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