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2sa mutant constructs  (Addgene inc)


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    Addgene inc 2sa mutant constructs
    Figure 4 Notch1 intracellular domain induces ubiquitination and degradation of the Snail protein. (A) Hep3B cells were transfected with Myc-Notch1 intracellular domain (NICD) and/or hemagglutinin (HA)-Snail, then treated with MG132 for 6 hours. Expression of Myc-NICD and HA- Snail was then analyzed by immunoblotting. (B) HA-Snail- or HA-Snail/Myc-NICD-transfected Hep3B cells were treated with 20 μM cycloheximide for different time intervals, then evaluated for Snail protein levels using immunoblot analysis. (C) 293T cells were transfected with Myc-NICD, SFB- Snail and/or HA-tagged ubiquitin, then treated with MG132 for 6 hours. Snail protein was immunoprecipitated using an anti-Flag antibody. Cell lysates were analyzed by immunoblotting with the indicated antibodies. (D) Hep3B cells were transfected with Myc-NICD, HA-Snail and MDM2; coimmunoprecipitated using anti-Myc and anti-MDM2 antibodies, respectively; and evaluated by immunoblot analysis. Immunoglobulin G served as a negative control. (E) Hep3B cells were transfected with Myc-NICD, HA-Snail and/or MDM2 siRNA and evaluated for the expression of Myc- NICD, HA-Snail and MDM2 using immunoblot analysis. (F) Hep3B cells were transfected with Myc-NICD and/or Flag-Snail wild-type (WT) or <t>2SA</t> mutant. The expression of Myc-NICD and Flag-Snail was then analyzed by immunoblotting. b-actin was used as an internal control.
    2sa Mutant Constructs, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/2sa+mutant+construct/pm22128911-182-3-9?v=Addgene+inc
    Average 93 stars, based on 2 article reviews
    2sa mutant constructs - by Bioz Stars, 2026-08
    93/100 stars

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    1) Product Images from "Notch1 binds and induces degradation of Snail in hepatocellular carcinoma."

    Article Title: Notch1 binds and induces degradation of Snail in hepatocellular carcinoma.

    Journal: BMC biology

    doi: 10.1186/1741-7007-9-83

    Figure 4 Notch1 intracellular domain induces ubiquitination and degradation of the Snail protein. (A) Hep3B cells were transfected with Myc-Notch1 intracellular domain (NICD) and/or hemagglutinin (HA)-Snail, then treated with MG132 for 6 hours. Expression of Myc-NICD and HA- Snail was then analyzed by immunoblotting. (B) HA-Snail- or HA-Snail/Myc-NICD-transfected Hep3B cells were treated with 20 μM cycloheximide for different time intervals, then evaluated for Snail protein levels using immunoblot analysis. (C) 293T cells were transfected with Myc-NICD, SFB- Snail and/or HA-tagged ubiquitin, then treated with MG132 for 6 hours. Snail protein was immunoprecipitated using an anti-Flag antibody. Cell lysates were analyzed by immunoblotting with the indicated antibodies. (D) Hep3B cells were transfected with Myc-NICD, HA-Snail and MDM2; coimmunoprecipitated using anti-Myc and anti-MDM2 antibodies, respectively; and evaluated by immunoblot analysis. Immunoglobulin G served as a negative control. (E) Hep3B cells were transfected with Myc-NICD, HA-Snail and/or MDM2 siRNA and evaluated for the expression of Myc- NICD, HA-Snail and MDM2 using immunoblot analysis. (F) Hep3B cells were transfected with Myc-NICD and/or Flag-Snail wild-type (WT) or 2SA mutant. The expression of Myc-NICD and Flag-Snail was then analyzed by immunoblotting. b-actin was used as an internal control.
    Figure Legend Snippet: Figure 4 Notch1 intracellular domain induces ubiquitination and degradation of the Snail protein. (A) Hep3B cells were transfected with Myc-Notch1 intracellular domain (NICD) and/or hemagglutinin (HA)-Snail, then treated with MG132 for 6 hours. Expression of Myc-NICD and HA- Snail was then analyzed by immunoblotting. (B) HA-Snail- or HA-Snail/Myc-NICD-transfected Hep3B cells were treated with 20 μM cycloheximide for different time intervals, then evaluated for Snail protein levels using immunoblot analysis. (C) 293T cells were transfected with Myc-NICD, SFB- Snail and/or HA-tagged ubiquitin, then treated with MG132 for 6 hours. Snail protein was immunoprecipitated using an anti-Flag antibody. Cell lysates were analyzed by immunoblotting with the indicated antibodies. (D) Hep3B cells were transfected with Myc-NICD, HA-Snail and MDM2; coimmunoprecipitated using anti-Myc and anti-MDM2 antibodies, respectively; and evaluated by immunoblot analysis. Immunoglobulin G served as a negative control. (E) Hep3B cells were transfected with Myc-NICD, HA-Snail and/or MDM2 siRNA and evaluated for the expression of Myc- NICD, HA-Snail and MDM2 using immunoblot analysis. (F) Hep3B cells were transfected with Myc-NICD and/or Flag-Snail wild-type (WT) or 2SA mutant. The expression of Myc-NICD and Flag-Snail was then analyzed by immunoblotting. b-actin was used as an internal control.

    Techniques Used: Ubiquitin Proteomics, Transfection, Expressing, Western Blot, Immunoprecipitation, Negative Control, Mutagenesis, Control



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    2sa mutant constructs  (Addgene inc)
    93
    Addgene inc 2sa mutant constructs
    Figure 4 Notch1 intracellular domain induces ubiquitination and degradation of the Snail protein. (A) Hep3B cells were transfected with Myc-Notch1 intracellular domain (NICD) and/or hemagglutinin (HA)-Snail, then treated with MG132 for 6 hours. Expression of Myc-NICD and HA- Snail was then analyzed by immunoblotting. (B) HA-Snail- or HA-Snail/Myc-NICD-transfected Hep3B cells were treated with 20 μM cycloheximide for different time intervals, then evaluated for Snail protein levels using immunoblot analysis. (C) 293T cells were transfected with Myc-NICD, SFB- Snail and/or HA-tagged ubiquitin, then treated with MG132 for 6 hours. Snail protein was immunoprecipitated using an anti-Flag antibody. Cell lysates were analyzed by immunoblotting with the indicated antibodies. (D) Hep3B cells were transfected with Myc-NICD, HA-Snail and MDM2; coimmunoprecipitated using anti-Myc and anti-MDM2 antibodies, respectively; and evaluated by immunoblot analysis. Immunoglobulin G served as a negative control. (E) Hep3B cells were transfected with Myc-NICD, HA-Snail and/or MDM2 siRNA and evaluated for the expression of Myc- NICD, HA-Snail and MDM2 using immunoblot analysis. (F) Hep3B cells were transfected with Myc-NICD and/or Flag-Snail wild-type (WT) or <t>2SA</t> mutant. The expression of Myc-NICD and Flag-Snail was then analyzed by immunoblotting. b-actin was used as an internal control.
    2sa Mutant Constructs, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/2sa+mutant+construct/pm22128911-182-3-9?v=Addgene+inc
    Average 93 stars, based on 1 article reviews
    2sa mutant constructs - by Bioz Stars, 2026-08
    93/100 stars
    		  Buy from Supplier

    Image Search Results


    Figure 4 Notch1 intracellular domain induces ubiquitination and degradation of the Snail protein. (A) Hep3B cells were transfected with Myc-Notch1 intracellular domain (NICD) and/or hemagglutinin (HA)-Snail, then treated with MG132 for 6 hours. Expression of Myc-NICD and HA- Snail was then analyzed by immunoblotting. (B) HA-Snail- or HA-Snail/Myc-NICD-transfected Hep3B cells were treated with 20 μM cycloheximide for different time intervals, then evaluated for Snail protein levels using immunoblot analysis. (C) 293T cells were transfected with Myc-NICD, SFB- Snail and/or HA-tagged ubiquitin, then treated with MG132 for 6 hours. Snail protein was immunoprecipitated using an anti-Flag antibody. Cell lysates were analyzed by immunoblotting with the indicated antibodies. (D) Hep3B cells were transfected with Myc-NICD, HA-Snail and MDM2; coimmunoprecipitated using anti-Myc and anti-MDM2 antibodies, respectively; and evaluated by immunoblot analysis. Immunoglobulin G served as a negative control. (E) Hep3B cells were transfected with Myc-NICD, HA-Snail and/or MDM2 siRNA and evaluated for the expression of Myc- NICD, HA-Snail and MDM2 using immunoblot analysis. (F) Hep3B cells were transfected with Myc-NICD and/or Flag-Snail wild-type (WT) or 2SA mutant. The expression of Myc-NICD and Flag-Snail was then analyzed by immunoblotting. b-actin was used as an internal control.

    Journal: BMC biology

    Article Title: Notch1 binds and induces degradation of Snail in hepatocellular carcinoma.

    doi: 10.1186/1741-7007-9-83

    Figure Lengend Snippet: Figure 4 Notch1 intracellular domain induces ubiquitination and degradation of the Snail protein. (A) Hep3B cells were transfected with Myc-Notch1 intracellular domain (NICD) and/or hemagglutinin (HA)-Snail, then treated with MG132 for 6 hours. Expression of Myc-NICD and HA- Snail was then analyzed by immunoblotting. (B) HA-Snail- or HA-Snail/Myc-NICD-transfected Hep3B cells were treated with 20 μM cycloheximide for different time intervals, then evaluated for Snail protein levels using immunoblot analysis. (C) 293T cells were transfected with Myc-NICD, SFB- Snail and/or HA-tagged ubiquitin, then treated with MG132 for 6 hours. Snail protein was immunoprecipitated using an anti-Flag antibody. Cell lysates were analyzed by immunoblotting with the indicated antibodies. (D) Hep3B cells were transfected with Myc-NICD, HA-Snail and MDM2; coimmunoprecipitated using anti-Myc and anti-MDM2 antibodies, respectively; and evaluated by immunoblot analysis. Immunoglobulin G served as a negative control. (E) Hep3B cells were transfected with Myc-NICD, HA-Snail and/or MDM2 siRNA and evaluated for the expression of Myc- NICD, HA-Snail and MDM2 using immunoblot analysis. (F) Hep3B cells were transfected with Myc-NICD and/or Flag-Snail wild-type (WT) or 2SA mutant. The expression of Myc-NICD and Flag-Snail was then analyzed by immunoblotting. b-actin was used as an internal control.

    Article Snippet: Flag-Snail WT and 2SA mutant constructs were obtained from Addgene, Inc (Cambridge, MA, USA).

    Techniques: Ubiquitin Proteomics, Transfection, Expressing, Western Blot, Immunoprecipitation, Negative Control, Mutagenesis, Control