glosensor camp-biosensor cdna Search Results


glosensor camp-biosensor cdna  (Promega)
90
Promega glosensor camp-biosensor cdna
RGS21 binds gustducin-α and reduces bitterant signaling in a bitterant-responsive cultured cell line. (A) COS7 cell monolayers were transiently transfected with plasmid DNA encoding gustducin-α tagged with an HA epitope (“HA-Gα gust”). Forty-eight hours post transfection, cell monolayers were lysed in buffer containing GDP, Mg2+ and AlF4– (“AMF”) or GDP alone, as indicated. Clarified cell lysates were incubated with 10 μg of purified His6-RGS21 protein at 4°C overnight with NTA-agarose. Samples were centrifuged, agarose beads washed four times in lysis buffer, and precipitated proteins (“P”:) eluted by boiling for 5 minutes in loading buffer prior to being resolved by SDS–PAGE electrophoresis on the basis of molecular weight (“kDa” = kiloDalton), transferred to a nitrocellulose membrane, and detected using anti-HA antibody immunoblotting (“IB”:) and chemiluminescence. (B) Overexpression of wild-type RGS21, but not a loss-of-function, point mutant RGS21, leads to inhibition of bitterant signaling-induced reduction of <t>cAMP</t> levels. Cultures of the 16HBE cell line were transiently transfected with <t>GloSensor</t> cAMP biosensor <t>cDNA</t> and expression plasmids containing open reading frames for wild-type (“wt”) RGS21, or Arg-126-to-Glu point mutated (“R>E”) RGS21, or empty pcDNA3.1 vector, as indicated. Inhibition of forskolin-stimulated cAMP production (“100% maximum”; dotted line) by treatment with indicated concentration of the bitterant denatonium benzoate (i.e. EC50 previously established as per Supplementary Fig. S1) was determined 24 hr post-transfection by detection of GloSensor-dependent luminescence. Asterisks denote statistically significant differences as determined by one-way ANOVA with Bonferroni’s post-test: **, P < 0.01; ns, not significant (P >> 0.05).
Glosensor Camp Biosensor Cdna, supplied by Promega, 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/result/glosensor camp-biosensor cdna/product/Promega
Average 90 stars, based on 1 article reviews
glosensor camp-biosensor cdna - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier

Image Search Results


RGS21 binds gustducin-α and reduces bitterant signaling in a bitterant-responsive cultured cell line. (A) COS7 cell monolayers were transiently transfected with plasmid DNA encoding gustducin-α tagged with an HA epitope (“HA-Gα gust”). Forty-eight hours post transfection, cell monolayers were lysed in buffer containing GDP, Mg2+ and AlF4– (“AMF”) or GDP alone, as indicated. Clarified cell lysates were incubated with 10 μg of purified His6-RGS21 protein at 4°C overnight with NTA-agarose. Samples were centrifuged, agarose beads washed four times in lysis buffer, and precipitated proteins (“P”:) eluted by boiling for 5 minutes in loading buffer prior to being resolved by SDS–PAGE electrophoresis on the basis of molecular weight (“kDa” = kiloDalton), transferred to a nitrocellulose membrane, and detected using anti-HA antibody immunoblotting (“IB”:) and chemiluminescence. (B) Overexpression of wild-type RGS21, but not a loss-of-function, point mutant RGS21, leads to inhibition of bitterant signaling-induced reduction of cAMP levels. Cultures of the 16HBE cell line were transiently transfected with GloSensor cAMP biosensor cDNA and expression plasmids containing open reading frames for wild-type (“wt”) RGS21, or Arg-126-to-Glu point mutated (“R>E”) RGS21, or empty pcDNA3.1 vector, as indicated. Inhibition of forskolin-stimulated cAMP production (“100% maximum”; dotted line) by treatment with indicated concentration of the bitterant denatonium benzoate (i.e. EC50 previously established as per Supplementary Fig. S1) was determined 24 hr post-transfection by detection of GloSensor-dependent luminescence. Asterisks denote statistically significant differences as determined by one-way ANOVA with Bonferroni’s post-test: **, P < 0.01; ns, not significant (P >> 0.05).

Journal: Chemical Senses

Article Title: The stability of tastant detection by mouse lingual chemosensory tissue requires Regulator of G protein Signaling-21 (RGS21)

doi: 10.1093/chemse/bjab048

Figure Lengend Snippet: RGS21 binds gustducin-α and reduces bitterant signaling in a bitterant-responsive cultured cell line. (A) COS7 cell monolayers were transiently transfected with plasmid DNA encoding gustducin-α tagged with an HA epitope (“HA-Gα gust”). Forty-eight hours post transfection, cell monolayers were lysed in buffer containing GDP, Mg2+ and AlF4– (“AMF”) or GDP alone, as indicated. Clarified cell lysates were incubated with 10 μg of purified His6-RGS21 protein at 4°C overnight with NTA-agarose. Samples were centrifuged, agarose beads washed four times in lysis buffer, and precipitated proteins (“P”:) eluted by boiling for 5 minutes in loading buffer prior to being resolved by SDS–PAGE electrophoresis on the basis of molecular weight (“kDa” = kiloDalton), transferred to a nitrocellulose membrane, and detected using anti-HA antibody immunoblotting (“IB”:) and chemiluminescence. (B) Overexpression of wild-type RGS21, but not a loss-of-function, point mutant RGS21, leads to inhibition of bitterant signaling-induced reduction of cAMP levels. Cultures of the 16HBE cell line were transiently transfected with GloSensor cAMP biosensor cDNA and expression plasmids containing open reading frames for wild-type (“wt”) RGS21, or Arg-126-to-Glu point mutated (“R>E”) RGS21, or empty pcDNA3.1 vector, as indicated. Inhibition of forskolin-stimulated cAMP production (“100% maximum”; dotted line) by treatment with indicated concentration of the bitterant denatonium benzoate (i.e. EC50 previously established as per Supplementary Fig. S1) was determined 24 hr post-transfection by detection of GloSensor-dependent luminescence. Asterisks denote statistically significant differences as determined by one-way ANOVA with Bonferroni’s post-test: **, P < 0.01; ns, not significant (P >> 0.05).

Article Snippet: Briefly, monolayer cultures of the 16HBE cell line were transiently cotransfected using FuGENE6 (Promega) with the GloSensor cAMP-biosensor cDNA (Promega) and pcDNA3.1-based expression plasmids encoding either wild-type RGS21 open-reading frame, or a GAP-dead, loss-of-function point-mutant version (R126E a.k.a.

Techniques: Cell Culture, Transfection, Plasmid Preparation, Incubation, Purification, Lysis, SDS Page, Electrophoresis, Molecular Weight, Western Blot, Over Expression, Mutagenesis, Inhibition, Expressing, Concentration Assay