Antibody-based Protein Detection Protocol

The following primary antibodies were used: Rabbit anti-SPBP antibody [27] (link), monoclonal anti-p62 antibody (BD Transduction Laboratories), rabbit anti-NRF2 antibody (Santa Cruz, SC-13032, Abcam ab62352), rabbit anti-LC3B antibody (SIGMA, L7543), rabbit anti-KEAP1 antibody (ProteinTech, 10503-2-AP), mouse anti-Flag antibody (Stratagene, 200471), rabbit anti-actin antibodies (Sigma, A 2066). Secondary antibodies used were: HRP-conjugated goat anti-rabbit IgG and anti-mouse IgG antibodies (BD Bioscience Pharmingen). AlexaFluor 568 and AlexaFluor 548 conjugated goat anti mouse and anti-rabbit IgG (Molecular Probes). Sulforaphane (S 4441), Bafilomycin A1 (B 1793) and MG 132 (C 2211) were purchased from Sigma-Aldrich.

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Darvekar S.R., Elvenes J., Brenne H.B., Johansen T, & Sjøttem E. (2014). SPBP Is a Sulforaphane Induced Transcriptional Coactivator of NRF2 Regulating Expression of the Autophagy Receptor p62/SQSTM1. PLoS ONE, 9(1), e85262.

Publication 2014

rabbit anti-NRF2 antibody rabbit anti-LC3B antibody rabbit anti-actin antibodies HRP-conjugated goat anti-rabbit IgG Bafilomycin A1 (B 1793) MG 132 (C 2211)

Actin Anti antibodies Anti antibody Anti igg Antibodies B 1793 Bafilomycin a1 Goat Keap1 Mg 132 Molecular probes Monoclonal antibody Mouse Nrf2 Rabbit Sulforaphane

Corresponding Organization :

Other organizations : UiT The Arctic University of Norway

Top 5 similar protocols

Variable analysis

independent variables

Sulforaphane (S 4441)
Bafilomycin A1 (B 1793)
MG 132 (C 2211)

dependent variables

KEAP1

control variables

Actin

positive controls

Rabbit anti-SPBP antibody [27]
Monoclonal anti-p62 antibody (BD Transduction Laboratories)
Rabbit anti-NRF2 antibody (Santa Cruz, SC-13032, Abcam ab62352)
Rabbit anti-LC3B antibody (SIGMA, L7543)
Rabbit anti-KEAP1 antibody (ProteinTech, 10503-2-AP)
Mouse anti-Flag antibody (Stratagene, 200471)

negative controls

Rabbit anti-actin antibodies (Sigma, A 2066)

Annotations

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