nu-879 Search Results


n 6 phenyl atp  (Jena Bioscience)
90
Jena Bioscience n 6 phenyl atp
(A) In vitro kinase assay with the purified analog-sensitive (AS) variant of CaMKIIβ. CaMKII activity against a protein substrate (Syntide 2, fused to GST) was measured as a function of calmodulin concentration. Direct phosphorylation of the substrate by the analog-sensitive F89G variant was measured via 32 P incorporation. Samples were separated on an SDS-PAGE and detected using autoradiography. (B) Quantification of A, normalized to the maximum activity at 400 nM calmodulin. Data for the wt variant of CaMKIIβΔ13 and CaMKIIβΔ13,16 are taken from . Error bars indicate standard deviation (n=3). (C) Inhibition of the AS variant of CaMKIIβ by various <t>N</t> <t>6</t> -modified <t>ATP</t> analogs. An in vitro kinase assay was performed at an optimal calmodulin concentration and supplemented with different non-radioactive ATP analogs. Enzymatic activity was measured as described for A. (D) Quantification of C, normalized to the non-inhibited signal (n=3). Note that the wt enzyme is not inhibited by N 6 -modified ATP analogs, whereas the AS variant is inhibited. (E) Western blot showing the labeling efficiency in permeabilized cells. N2a cells overexpressing Twin-Strep-CaMKIIIβΔ13,16-F89G were collected and permeabilized with nOG (n-octyl-β-D-glucopyranoside) or Tween-20 as indicated, or lysed by brief sonication. Reactions were performed with N 6 -benzyl-ATPγS in the presence/ absence of stimulating conditions (calmodulin/Ca 2+ ) and/or an external substrate (Syntide 2, linked to GST). Samples were alkylated, run on an SDS-PAGE and analyzed via western blot with a thiophosphate ester-specific antibody. (F) Correlation matrix of the substrate spectra of different CaMKIIβ isoforms, as determined by an analog-sensitive kinase assay. The analysis was restricted to CaMKIIβ-specific targets. Additionally, CaMKIIβ autophosphorylation targets were removed from the analysis. A Person correlation coefficient was calculated based on the intensity values of individual phosphorylation sites.
N 6 Phenyl Atp, supplied by Jena Bioscience, 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/n 6 phenyl atp/product/Jena Bioscience
Average 90 stars, based on 1 article reviews
n 6 phenyl atp - by Bioz Stars, 2026-02
90/100 stars
  Buy from Supplier

Image Search Results


(A) In vitro kinase assay with the purified analog-sensitive (AS) variant of CaMKIIβ. CaMKII activity against a protein substrate (Syntide 2, fused to GST) was measured as a function of calmodulin concentration. Direct phosphorylation of the substrate by the analog-sensitive F89G variant was measured via 32 P incorporation. Samples were separated on an SDS-PAGE and detected using autoradiography. (B) Quantification of A, normalized to the maximum activity at 400 nM calmodulin. Data for the wt variant of CaMKIIβΔ13 and CaMKIIβΔ13,16 are taken from . Error bars indicate standard deviation (n=3). (C) Inhibition of the AS variant of CaMKIIβ by various N 6 -modified ATP analogs. An in vitro kinase assay was performed at an optimal calmodulin concentration and supplemented with different non-radioactive ATP analogs. Enzymatic activity was measured as described for A. (D) Quantification of C, normalized to the non-inhibited signal (n=3). Note that the wt enzyme is not inhibited by N 6 -modified ATP analogs, whereas the AS variant is inhibited. (E) Western blot showing the labeling efficiency in permeabilized cells. N2a cells overexpressing Twin-Strep-CaMKIIIβΔ13,16-F89G were collected and permeabilized with nOG (n-octyl-β-D-glucopyranoside) or Tween-20 as indicated, or lysed by brief sonication. Reactions were performed with N 6 -benzyl-ATPγS in the presence/ absence of stimulating conditions (calmodulin/Ca 2+ ) and/or an external substrate (Syntide 2, linked to GST). Samples were alkylated, run on an SDS-PAGE and analyzed via western blot with a thiophosphate ester-specific antibody. (F) Correlation matrix of the substrate spectra of different CaMKIIβ isoforms, as determined by an analog-sensitive kinase assay. The analysis was restricted to CaMKIIβ-specific targets. Additionally, CaMKIIβ autophosphorylation targets were removed from the analysis. A Person correlation coefficient was calculated based on the intensity values of individual phosphorylation sites.

Journal: bioRxiv

Article Title: Branch point evolution controls species-specific alternative splicing and regulates long term potentiation

doi: 10.1101/2022.09.09.507289

Figure Lengend Snippet: (A) In vitro kinase assay with the purified analog-sensitive (AS) variant of CaMKIIβ. CaMKII activity against a protein substrate (Syntide 2, fused to GST) was measured as a function of calmodulin concentration. Direct phosphorylation of the substrate by the analog-sensitive F89G variant was measured via 32 P incorporation. Samples were separated on an SDS-PAGE and detected using autoradiography. (B) Quantification of A, normalized to the maximum activity at 400 nM calmodulin. Data for the wt variant of CaMKIIβΔ13 and CaMKIIβΔ13,16 are taken from . Error bars indicate standard deviation (n=3). (C) Inhibition of the AS variant of CaMKIIβ by various N 6 -modified ATP analogs. An in vitro kinase assay was performed at an optimal calmodulin concentration and supplemented with different non-radioactive ATP analogs. Enzymatic activity was measured as described for A. (D) Quantification of C, normalized to the non-inhibited signal (n=3). Note that the wt enzyme is not inhibited by N 6 -modified ATP analogs, whereas the AS variant is inhibited. (E) Western blot showing the labeling efficiency in permeabilized cells. N2a cells overexpressing Twin-Strep-CaMKIIIβΔ13,16-F89G were collected and permeabilized with nOG (n-octyl-β-D-glucopyranoside) or Tween-20 as indicated, or lysed by brief sonication. Reactions were performed with N 6 -benzyl-ATPγS in the presence/ absence of stimulating conditions (calmodulin/Ca 2+ ) and/or an external substrate (Syntide 2, linked to GST). Samples were alkylated, run on an SDS-PAGE and analyzed via western blot with a thiophosphate ester-specific antibody. (F) Correlation matrix of the substrate spectra of different CaMKIIβ isoforms, as determined by an analog-sensitive kinase assay. The analysis was restricted to CaMKIIβ-specific targets. Additionally, CaMKIIβ autophosphorylation targets were removed from the analysis. A Person correlation coefficient was calculated based on the intensity values of individual phosphorylation sites.

Article Snippet: The reaction mixture contained varying concentrations of non-radioactive ATP (0-1 mM) or 0.5 mM of one of the following non-radioactive ATP analogs: N 6 -methyl-ATP, N 6 -etheno-ATP, N 6 -phenyl-ATP, N 6 -benzyl-ATP (Jena Bioscience).

Techniques: In Vitro, Kinase Assay, Purification, Variant Assay, Activity Assay, Concentration Assay, SDS Page, Autoradiography, Standard Deviation, Inhibition, Modification, Western Blot, Labeling, Sonication