Immunofluorescence Indirect Direct proof of antigens Ag proof
- Slides: 10
Immunofluorescence Indirect Direct proof of antigens (Ag) proof of antibodies (Ab) antibody against Ab+ colour Ab+ fluorescent colour (factory-made) serum (Ag) serum (Ig. G) Ag (f-m)
Immunofluorescence Task 1
ELISA I. (Enzyme-Linked Immunosorbent Assay) ® Is used in a proof of antigens and antibodies
ELISA: proof of antigen colour change + Substrate Ab marked with enzyme Serum with Ag Serum without Ag Ab bounded in well
ELISA II. – proof of antibodies ® We detect classes of antibodies – Ig. M, Ig. A, Ig. G ® !!! Ig. M and Ig. A demonstrate an acute infection ® Ig. G generally = infection in past, but !! ® exist so-called low avidity Ig. G – binding of these antibodies in reaction is not solid, it could be damaged by urea (they are present in earlier phase of infection)
ELISA: proof of antibodies general + Substrate Ab against antibody marked with enzyme Serum with Ab Serum without Ab Ag on the bottom
ELISA: proof of antibody classes – binding (capture) + - Substrate Ab against antibody markedwith enzyme Ag (factorymade) Serum without Ab/with other Ab than searching Serum with Ab (Ig. M/Ig. G) Ab against Ig. M//Ig. G bounds all Ig. M/Ig. G in sample
Westernblot (proof of Ab) Prepared Ag splitted by Ag DDS Cutting Strips gel. ELF O parted Suck to cellulose membran e
Westernblot + serum with antibodies (Ab) Firm-made strip with Ag (Bmp. A, Osp. C, p 39 etc. ) Binding Ab to Ag Osp. C sampl pattern e 2 lines similary with pattern = At least presence of antibodies Exception: ab against Osp. C u Ig. M line ab against vls. E u Ig. G enough 1 is
Pattern task no. 5 Osp. C is high specific Ig. M against Osp. C present = acute infection, we don´t need to find Ig. M against other antigens