Western Blot Analysis of Protein Targets

Immunoblotting was performed as previously described (13 (link)). The primary antibodies used were: (i) anti-TEG-1 (N-terminal) antibodies at 1:1000 (13 (link)); (ii) anti-VIG-1 antibodies at 1:30 000; (iii) anti-ALG-1 antibodies (Thermo Scientific) at 1:2500; (iv) anti-AGO2 antibodies (Cell Signaling) at 1:1000; (v) anti-GFP antibodies (Rockland) at 1:2000; (vi) anti-SERBP1 antibodies (anti-PAI-RBP1, Abnova) at 1:1000; (vii) anti-actin antibodies (Sigma) at 1:1000; (viii) rabbit anti-ubiquitin antibodies (Sigma) at 1:100 and (ix) anti-paramyosin antibodies (MH16, hybridoma cell line from Developmental Studies Hybridoma Bank, University of Iowa) tissue culture supernatant. Immunoblots were developed with SuperSignal WestPico substrate (Thermo Scientific) or Amersham ECL Select Western Blotting Detection Reagent (GE Healthcare). Blots were exposed to Kodak BioMax MR film.

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Wang C., Gupta P., Fressigne L., Bossé G.D., Wang X., Simard M.J, & Hansen D. (2016). TEG-1 CD2BP2 controls miRNA levels by regulating miRISC stability in C. elegans and human cells. Nucleic Acids Research, 45(3), 1488-1500.

Publication 2016

anti-AGO2 antibodies anti-GFP antibodies anti-actin antibodies SuperSignal WestPico substrate Amersham ECL Select Western Blotting Detection Reagent BioMax MR film

Actin Ago2 Anti antibodies Antibodies Cell line Hybridoma Immunoblots Paramyosin Rabbit Tissue Ubiquitin

Corresponding Organization :

Other organizations : University of Calgary, Hôtel-Dieu de Québec, Centre hospitalier universitaire de Québec, Université Laval

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Variable analysis

independent variables

Primary antibodies used:
(i) anti-TEG-1 (N-terminal) antibodies at 1:1000
(ii) anti-VIG-1 antibodies at 1:30 000
(iii) anti-ALG-1 antibodies (Thermo Scientific) at 1:2500
(iv) anti-AGO2 antibodies (Cell Signaling) at 1:1000
(v) anti-GFP antibodies (Rockland) at 1:2000
(vi) anti-SERBP1 antibodies (anti-PAI-RBP1, Abnova) at 1:1000
(vii) anti-actin antibodies (Sigma) at 1:1000
(viii) rabbit anti-ubiquitin antibodies (Sigma) at 1:100
(ix) anti-paramyosin antibodies (MH16, hybridoma cell line from Developmental Studies Hybridoma Bank, University of Iowa) tissue culture supernatant

dependent variables

Protein levels detected by immunoblotting

control variables

Immunoblotting was performed as previously described (13 (link)).

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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