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pcmv3 ha keap1  (Sino Biological)


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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 3 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
    Average 91 stars, based on 3 article reviews
    pcmv3 ha keap1 - by Bioz Stars, 2026-07
    91/100 stars

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    1) Product Images from "Chemoproteomic profiling reveals cellular targets of nitro-fatty acids"

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

    Journal: Redox Biology

    doi: 10.1016/j.redox.2021.102126

    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.
    Figure Legend 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.

    Techniques Used: Fluorescence, Labeling, Molecular Weight



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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 <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  (Sino Biological)
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    (A) Workflow for in-gel fluorescence detection of nitro-alkylated proteins using the clickable 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 by alk-9-NO 2 -OA. (D) Labeling of wildtype (wt) KEAP1 and its mutants in HEK293FT cells at 2.5 μM alk-9-NO 2 -OA. 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. 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: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcmv3+ha+keap1/bio_rxiv__2021__07__12__451990-43-4-5?v=Sino+Biological
    Average 90 stars, based on 1 article reviews
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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

    (A) Workflow for in-gel fluorescence detection of nitro-alkylated proteins using the clickable 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 by alk-9-NO 2 -OA. (D) Labeling of wildtype (wt) KEAP1 and its mutants in HEK293FT cells at 2.5 μM alk-9-NO 2 -OA. 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. 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: bioRxiv

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

    doi: 10.1101/2021.07.12.451990

    Figure Lengend Snippet: (A) Workflow for in-gel fluorescence detection of nitro-alkylated proteins using the clickable 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 by alk-9-NO 2 -OA. (D) Labeling of wildtype (wt) KEAP1 and its mutants in HEK293FT cells at 2.5 μM alk-9-NO 2 -OA. 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. 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) were made using the QuikChange Lightning Multi Site-Directed Mutagenesis Kit (Stratagene) and validated by Sanger sequencing.

    Techniques: Fluorescence, Labeling, Molecular Weight