Western Blot Analysis of TRPC3, Cadherin, and GAPDH

The procedures of Western blot were carried out as previously described [15 (link)]. Protein samples (20μg) were separated in an 8% DS-PAGE gel and transferred to a nitrocellulose membrane (#66485; PALL Life Sciences) via semi-dry transfer (350 mA, 90 min). Primary antibodies: Rabbit anti-TRPC3, 1:200, #ab 51560, Abcam; rabbit anti-Pan cadherin, 1:1000, ab6505, #ab16505, Abcam; mouse anti-GAPDH, 1:20,000, #2118S, Cell Signaling Technology. Densitometry was employed to quantify proteins of interest using the gel analyzer function of ImageJ (ver. 1.46a; National Institutes of Health, USA).

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Deurloo M.H., Turlova E., Chen W.L., Lin Y.W., Tam E., Tassew N.G., Wu M., Huang Y.C., Crawley J.N., Monnier P.P., Groffen A.J., Sun H.S., Osborne L.R, & Feng Z.P. (2018). Transcription Factor 2I Regulates Neuronal Development via TRPC3 in 7q11.23 Disorder Models. Molecular Neurobiology, 56(5), 3313-3325.

Publication 2018

ab 51560 ab6505 mouse anti-GAPDH

Antibodies Cadherin Densitometry Gapdh Membrane Mouse Nitrocellulose Proteins Rabbit Western blot

Corresponding Organization :

Other organizations : Canada Research Chairs, University of Toronto, Krembil Research Institute, Krembil Foundation, University of California, Davis, Amsterdam Neuroscience

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Variable analysis

independent variables

Protein sample type (e.g., different cell or tissue samples)

dependent variables

Protein expression levels of TRPC3 and Pan cadherin

control variables

Protein loading amount (20 μg)
Gel type (8% DS-PAGE)
Transfer method (semi-dry transfer, 350 mA, 90 min)
Primary antibodies (Rabbit anti-TRPC3, Rabbit anti-Pan cadherin, Mouse anti-GAPDH)

controls

Positive control: Pan cadherin (a housekeeping protein)
Negative control: Not explicitly mentioned

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