[7] Purification and assay of cAMP, cGMP, and cyclic nucleotide analogs in cells treated with cyclic nucleotide analogs
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Cited by (29)
Expression and distribution of cyclic GMP-dependent protein kinase-1 isoforms in human penile erectile tissue
2008, Journal of Sexual MedicineCitation Excerpt :To verify the localization of cGKI within the smooth musculature, double stainings using the monoclonal smooth muscle α-actin antibody (A5228, Sigma Chemical Co., St. Louis, MO, USA; 1:1000) [25] were carried out. Moreover, an anti-cGMP rabbit polyclonal antibody (Calbiochem, 1:500) [26] and the nitric oxide synthase III (eNOS) polyclonal antibody (Chemicon International, Temecula, CA, USA; 1:500) were used in order to show the colocalization of cGKI and its effector cGMP or cGKI and eNOS, respectively. The following Alexa Fluor (Molecular Probes Europe BV) secondary antibodies were used: #A11001, #A11008, #A11011, #A11055, and #A11057.
Immunohistochemical Distribution of Cyclic GMP-Dependent Protein Kinase-1 in Human Prostate Tissue
2007, European UrologyCitation Excerpt :To verify the localization of cGKI within the smooth musculature, double-staining using the monoclonal smooth muscle α-actin antibody (A5228, Sigma Chemical Co., St. Louis, MO, USA; 1:1000) [16] was carried out. In addition, an anti-cGMP rabbit polyclonal antibody (Calbiochem; 1:500) [17] was used to show the colocalization of cGKI and its effector protein cGMP. Fresh frozen tissue samples of the transition zone of human prostate tissue were sectioned into 20-μm slices and collected in Eppendorf tubes.
Ethanol stimulates ciliary beating by dual cyclic nucleotide kinase activation in bovine bronchial epithelial cells
2003, American Journal of PathologyCitation Excerpt :The papers were then washed five times manually for 1 minute each, rinsed 1 minute in EtOH, dried, and counted in nonaqueous scintillant.38 The assay for cGMP levels was performed similarly to that for cAMP.36 The PKG used was partially purified from bovine lung as previously described.39
A conserved serine juxtaposed to the pseudosubstrate site of type I cGMP-dependent protein kinase contributes strongly to autoinhibition and lower cGMP affinity
2002, Journal of Biological ChemistryCitation Excerpt :This value was within 1.5–2-fold of the value obtained from the cGMP kinase activation studies (Table I). As expected, the WT and S64A cGMP binding affinities were greater at 4 °C (34). Interestingly, the KD of either S64D or S64N was similar at 30 and 4 °C (2.0 ± 0.3 and 2.1 ± 0.4 nm; 1.7 ± 0.5 and 2.6 ± 1.1 nm, respectively).
In vitro PKA phosphorylation-mediated human PDE4A4 activation
2002, FEBS Letters