Immunofluorescence and Western Blotting Protocols

The following primary antibodies were used rabbit antiserum R18024 and R18611 [40] (link), rabbit anti-skeletal myosin heavy chain IgG (Santa Cruz Biotechnology), mouse anti-EB1 (BD Biosciences), -β-tubulin (Thermo Scientific), -vinculin (VII-F9, [60] (link)), -GM130 (BD biosciences), -FLAG-M2 antibody (Sigma-Aldrich), and rat anti-FLAG tag (Stratagene). Secondary antibodies used for immunofluorescence studies were highly cross-absorbed Alexa Fluor 488 and 568 anti-rabbit IgG, Alexa Fluor 488 and 568 anti-mouse, and Alexa Fluor 488 anti-rat, all from Invitrogen. For western blotting secondary antibodies were IR-Dye 800cw anti-rabbit IgG (LI-COR) and anti-mouse Alexa 680 (Invitrogen).

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Poliakova K., Adebola A., Leung C.L., Favre B., Liem R.K., Schepens I, & Borradori L. (2014). BPAG1a and b Associate with EB1 and EB3 and Modulate Vesicular Transport, Golgi Apparatus Structure, and Cell Migration in C2.7 Myoblasts. PLoS ONE, 9(9), e107535.

Publication 2014

mouse anti-EB1 -β-tubulin -vinculin -GM130 -FLAG-M2 antibody Alexa Fluor 488 anti-rat IR-Dye 800cw anti-rabbit IgG

Alexa fluor 488 Anti igg Antibodies Antibody Antiserum Immunofluorescence Mouse Myosin heavy chain Rabbit Skeletal Tubulin Vinculin

Corresponding Organization : Columbia University

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Variable analysis

independent variables

Rabbit antiserum R18024 and R18611
Rabbit anti-skeletal myosin heavy chain IgG (Santa Cruz Biotechnology)
Mouse anti-EB1 (BD Biosciences)
Mouse anti-β-tubulin (Thermo Scientific)
Mouse anti-vinculin (VII-F9)
Mouse anti-GM130 (BD biosciences)
Mouse anti-FLAG-M2 antibody (Sigma-Aldrich)
Rat anti-FLAG tag (Stratagene)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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