Immunostaining Protocols for α-Synuclein

The rat monoclonal antibody to human α-synuclein (15G7) was obtained from Alexis Biochemicals^{52 (link)}, the mouse monoclonal antibody to rodent α-synuclein (Syn-1) from BD Biosciences^{53 (link)}, the the goat polyclonal antibody to TOM20 (C-20) from Santa Cruz Biotechnology^{54 (link)}, the guinea pig polyclonal antibody to vesicular glutamate transporter 1 (VGLUT1) from EMD Millipore^{55 (link)} and the rabbit polyclonal antibodies to secretogranin II (K55101R) from Meridian Life Science^{56 (link)} and to actin (A2066) from Sigma^{57 (link)}. The H3C antibody to synuclein developed by J. George was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of NIH and maintained at the University of Iowa, Department of Biology, Iowa City, IA 52242. The anti-rat antibody conjugated to Alexa Fluor 488, anti-mouse antibody conjugated to Alexa Fluor 488, anti-goat antibody conjugated to Alexa Fluor 594 and anti-rabbit antibody conjugated to Alexa Fluor 594 were all obtained from Thermo Fisher Scientific. The anti-mouse antibody conjugated to Cy5 and anti-guinea pig antibody conjugated to Cy3 were obtained from Jackson ImmunoResearch. The anti-mouse antibody conjugated to IRDye800 and anti-rabbit antibody conjugated to IRDye680 were obtained from Rockland Immunochemicals.

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Logan T., Bendor J., Toupin C., Thorn K, & Edwards R.H. (2017). α-Synuclein Promotes Dilation of the Exocytotic Fusion Pore. Nature neuroscience, 20(5), 681-689.

Publication 2017

Syn-1) TOM20 (C-20) secretogranin II (K55101R) to actin (A2066) anti-rat antibody conjugated to Alexa Fluor 488 anti-mouse antibody conjugated to Alexa Fluor 488 anti-goat antibody conjugated to Alexa Fluor 594 anti-rabbit antibody conjugated to Alexa Fluor 594 anti-mouse antibody conjugated to Cy5 anti-guinea pig antibody conjugated to Cy3 anti-mouse antibody conjugated to IRDye800 anti-rabbit antibody conjugated to IRDye680

Actin Alexa 594 Alexa fluor 488 Anti antibody Antibodies Antibody Goat Guinea pig Human Hybridoma Irdye800 Meridian Monoclonal antibody Mouse Rabbit Rodent Secretogranin ii Synuclein Vesicular glutamate transporter 1 α synuclein

Corresponding Organization :

Other organizations : University of California, San Francisco

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Variable analysis

independent variables

Antibody type (rat monoclonal, mouse monoclonal, goat polyclonal, guinea pig polyclonal, rabbit polyclonal)

dependent variables

Not explicitly mentioned

control variables

Antibody sources (Alexis Biochemicals, BD Biosciences, Santa Cruz Biotechnology, EMD Millipore, Meridian Life Science, Sigma, Developmental Studies Hybridoma Bank)
Fluorescent dye-conjugated secondary antibodies (Alexa Fluor 488, Alexa Fluor 594, Cy5, Cy3, IRDye800, IRDye680)

controls

Positive control: H3C antibody to synuclein developed by J. George, obtained from the Developmental Studies Hybridoma Bank
Negative controls: Not explicitly mentioned

Annotations

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