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Sino Biological pcmv3 ha keap1
Fluorescent detection of nitro-alkylated proteins in living cells. (A) Workflow for in-gel fluorescence detection of nitro-alkylated proteins using the alk-9-NO 2 -OA probe 8 and azide-rhodamine. CuAAC, copper-catalyzed azide-alkyne cycloaddition. (B) Time-dependent labelling of endogenous proteins and overexpressed <t>HA-KEAP1</t> in HEK293FT cells at 2.5 μM alk-9-NO 2 -OA. (C) Dose-dependent labelling of endogenous proteins and overexpressed HA-KEAP1 in HEK293FT cells after treatment with indicated concentrations of alk-9-NO 2 -OA for 1 h. (D) Labeling of wildtype (wt) KEAP1 and its mutants in HEK293T cells at 2.5 μM alk-9-NO 2 -OA for 1 h. 3M and 7M refer to mutants with 3 and 7 cysteines mutated, respectively. (E) Fluorescent detection of nitro-alkylated proteins in different human cell lines treated with 10 μM alk-9-NO 2 -OA for 15 min. Anti-HA, anti-GAPDH, anti-tubulin blots and Coomassie stains act as loading controls for the accompanying fluorescence gels. Selected protein molecular weight markers are indicated for each gel.
Pcmv3 Ha Keap1, supplied by Sino Biological, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pcmv3+ha+keap1/pmc08441202-49-4-5?v=Sino+Biological
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pcmv3 ha keap1 - by Bioz Stars, 2026-07
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Fluorescent detection of nitro-alkylated proteins in living cells. (A) Workflow for in-gel fluorescence detection of nitro-alkylated proteins using the alk-9-NO 2 -OA probe 8 and azide-rhodamine. CuAAC, copper-catalyzed azide-alkyne cycloaddition. (B) Time-dependent labelling of endogenous proteins and overexpressed HA-KEAP1 in HEK293FT cells at 2.5 μM alk-9-NO 2 -OA. (C) Dose-dependent labelling of endogenous proteins and overexpressed HA-KEAP1 in HEK293FT cells after treatment with indicated concentrations of alk-9-NO 2 -OA for 1 h. (D) Labeling of wildtype (wt) KEAP1 and its mutants in HEK293T cells at 2.5 μM alk-9-NO 2 -OA for 1 h. 3M and 7M refer to mutants with 3 and 7 cysteines mutated, respectively. (E) Fluorescent detection of nitro-alkylated proteins in different human cell lines treated with 10 μM alk-9-NO 2 -OA for 15 min. Anti-HA, anti-GAPDH, anti-tubulin blots and Coomassie stains act as loading controls for the accompanying fluorescence gels. Selected protein molecular weight markers are indicated for each gel.

Journal: Redox Biology

Article Title: Chemoproteomic profiling reveals cellular targets of nitro-fatty acids

doi: 10.1016/j.redox.2021.102126

Figure Lengend Snippet: Fluorescent detection of nitro-alkylated proteins in living cells. (A) Workflow for in-gel fluorescence detection of nitro-alkylated proteins using the alk-9-NO 2 -OA probe 8 and azide-rhodamine. CuAAC, copper-catalyzed azide-alkyne cycloaddition. (B) Time-dependent labelling of endogenous proteins and overexpressed HA-KEAP1 in HEK293FT cells at 2.5 μM alk-9-NO 2 -OA. (C) Dose-dependent labelling of endogenous proteins and overexpressed HA-KEAP1 in HEK293FT cells after treatment with indicated concentrations of alk-9-NO 2 -OA for 1 h. (D) Labeling of wildtype (wt) KEAP1 and its mutants in HEK293T cells at 2.5 μM alk-9-NO 2 -OA for 1 h. 3M and 7M refer to mutants with 3 and 7 cysteines mutated, respectively. (E) Fluorescent detection of nitro-alkylated proteins in different human cell lines treated with 10 μM alk-9-NO 2 -OA for 15 min. Anti-HA, anti-GAPDH, anti-tubulin blots and Coomassie stains act as loading controls for the accompanying fluorescence gels. Selected protein molecular weight markers are indicated for each gel.

Article Snippet: The following mutations to pCMV3-HA-KEAP1 (Sino Biological, HG11981-NY) were made using the QuikChange Lightning Multi Site-Directed Mutagenesis Kit (Stratagene) and validated by Sanger sequencing ( ).

Techniques: Fluorescence, Labeling, Molecular Weight