%0 Figure %A J. Newman, Andrew %A Selkoe, Dennis %A Dettmer, Ulf %D 2013 %T In vitro incubation of lysates with 2 mM DSP followed by βME reduction allows optimal αSyn detection. %U https://plos.figshare.com/articles/figure/_In_vitro_incubation_of_lysates_with_2_mM_DSP_followed_by_ME_reduction_allows_optimal_Syn_detection_/858425 %R 10.1371/journal.pone.0081314.g003 %2 https://ndownloader.figshare.com/files/1290174 %K incubation %K lysates %K mm %K dsp %K followed %K allows %K optimal %X

A. HEL intact cells (in vivo) or lysates (in vitro) were incubated with DMSO only (-) as well as gradients of 0.5, 2.0 and 8.0 mM DSP and DTBP, respectively. Cytosols (post-100,000 g) were boiled in sample buffer/5% βME and blotted with αSyn antibody 2F12. Identical exposures of the same blot are shown; film was cut at dotted line. B. HEL cell cytosols (post-100,000 g) at three different protein concentrations (2.4, 3.3., 4.5 µg/µL) were incubated with DMSO only (-) as well as a gradient of 0.5, 2.0 and 8.0 mM DSP and DSG. Samples were normalized to 2.4 µg after quenching, then boiled in sample buffer plus 5% βME and blotted with αSyn antibodies Syn1 and 15G7 as well as an antibody to GAPDH. C. HEL cell cytosols (post-100,000 g) were treated with DMSO-only (-) or DSP, quenched, boiled in sample buffer plus 5% βME and analyzed by blotting for αSyn (mAb Syn1 and pAb C20), Calmodulin, DJ-1, Ran, 14-3-3 and β-actin.

%I PLOS ONE