Immunoprecipitation and Western Blotting

Immunoprecipitation was performed as we previously described 26 (link),27 (link). Cell lysates were immunoprecipitated with TMEM16A, p62, LC3B-I/II, bcl-2, Beclin-1 and VPS3 antibodies (those described for the western blotting protocol) using protein A/G agarose (sc-2003; Santa Cruz Biotechnology). The immunoprecipitates were washed with lysis buffer thrice and subjected to western blotting analysis. The bound proteins were immunoblotted with the indicated antibodies.

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Lv X.F., Zhang Y.J., Liu X., Zheng H.Q., Liu C.Z., Zeng X.L., Li X.Y., Lin X.C., Lin C.X., Ma M.M., Zhang F.R., Shang J.Y., Zhou J.G., Liang S.J, & Guan Y.Y. (2020). TMEM16A ameliorates vascular remodeling by suppressing autophagy via inhibiting Bcl-2-p62 complex formation. Theranostics, 10(9), 3980-3993.

Publication 2020

protein A/G agarose (sc-2003;

Agarose Antibodies Bcl 2 Beclin 1 Buffer Cell Immunoprecipitation Protein a Protein g Proteins Western blotting analysis

Corresponding Organization :

Other organizations : Sun Yat-sen University

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Variable analysis

independent variables

TMEM16A antibody
P62 antibody
LC3B-I/II antibody
Bcl-2 antibody
Beclin-1 antibody
VPS3 antibody

dependent variables

Bound proteins from immunoprecipitation

control variables

Protein A/G agarose (sc-2003; Santa Cruz Biotechnology)
Lysis buffer (used for washing the immunoprecipitates)

controls

Positive controls: Not specified
Negative controls: Not specified

Annotations

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