A novel enzyme-linked immunosorbent assay for determination of synaptophysin as compared with other quantification procedures.
A two-sided enzyme-linked immunosorbent assay (ELISA) has been established for reliable, specific and sensitive determination of synaptophysin (SYN), an intrinsic membrane protein of the small synaptic vesicles. This ELISA used a highly specific monoclonal antibody (SY 38) as capture reagent and a specific SYN antiserum in combination with a secondary peroxidase-conjugated antibody for detection. Calibration was carried out with immunoaffinity-purified SYN from porcine cortex. The sensitivity was found to be improved substantially when the ELISA was compared with previously used dot-immunobinding assays. This ELISA allowed rapid and reliable determination of SYN from detergent lysed homogenates, partially and highly purified preparations of rat, porcine and human brain. SYN was determined in highly purified small synaptic vesicles, and it was calculated to be 5.8% of total detergent solubilized protein.
Schlaf
G
G
Neurological Department of the University, Göttingen, Germany.
Salje
C
C
Poethke
R
R
Felgenhauer
K
K
Mäder
M
M
D000818:Animals; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D015151:Immunoblotting; D008854:Microscopy, Electron; D011817:Rabbits; D051381:Rats; D011917:Rats, Inbred Lew; D013552:Swine; D013572:Synaptic Vesicles / Q000737:chemistry / Q000648:ultrastructure; D016708:Synaptophysin / Q000032:analysis* / Q000302:isolation & purification
D016708:Synaptophysin