Citations Search

Notes: These authors investigated the mechanism of action of the migraine mediator calcitonin gene-related peptide (CGRP), which is known to sensitize the P2X₃ pain receptors to increase impulse flow to brain stem trigeminal nuclei. They showed that CAM KII inhibitors prevented CGRP-induced upregulation of P2X₃ mRNA using real-time RT-PCR, and then confirmed that active CAM KII was involved in the signaling mechanism by staining with Anti-ACTIVE^® CAM KII Antibodies. The immunoreactivity was upregulated by CGRP treatment of trigeminal neurons, and the distribution of the active CAM KII was localized to the membrane region. They then examined the mechanism of transcriptional activation of the P2X₃ pain receptor genes and showed that the transcription factor CREB, which is known to be dependent on CAM KII for activation and nuclear localization, was involved. The authors also investigated the role of BDNF in the process, because CGRP is known to promote BDNF expression by trigeminal neurons, and because BDNF is thought to be involved in pain processing. They used the BDNF E_max Immunoassay System to measure BDNF levels in culture media after application of CGRP to neuronal cell cultures, and demonstrated that there was a fourfold increase in BDNF after CGRP exposure. They also showed that BDNF promoted CAMKII and CREB activation. (3873)

Notes: Acidification can occur during synaptic nerve impulse transmission when the synaptic vesicles empty their contents into the synapse. The authors of this study investigated the role of protons in ion channel regulation at the synapse. They identified the acid-sensing ion channel 1a (ASIC1a) as a proton receptor, and they investigated its relationship to activation of the Ca²⁺/calmodulin-dependent protein kinase (CaM KII). Anti-ACTIVE^® CaM KII pAb was used for Western analysis of immunoprecipitated CaM KII from wildtype or ASIC1a-knockout mouse brains. (3549)

Notes: The authors of this study investigated the role of CaM KII in the induction of NMDA receptor-dependent sensitization of dorsal horn neurons. Ultrathin dorsal horn sections from capsaicin-injected and control rats were fixed and embedded and then incubated with Anti-ACTIVE^® CaM KII pAb. After washing, sections were incubated with goat anti-rabbit secondary antibody conjugated to 10nm colloidial gold. (3548)

Notes: Whole-cell extracts were prepared from rat ventricular myocytes treated with various concentrations of phenylephrine. Activation of CaMKII was assessed by Western analysis using Anti-ACTIVE^® CaMKII pAb. Anti-ACTIVE^® CaMKII pAb was also used to localize phosphorylated CaMKII by immunofluorescence microscopy of ventricular myocytes. Anti-ACTIVE^® CaMKII pAb and 15nm gold-conjugated goat anti-rabbit IgG secondary antibodies were used to perform immunoelectron microscopy of isolated myocytes to further determine the subcellular localization of activated CaMKII. (3435)

Notes: Anti-ACTIVE^® CaM KII pAb was used to detect phosphorylated CAM KII by immunohistochemistry and Western blot analysis in tissues isolated from transgenic mice. A 1:200 dilution of the Anti-ACTIVE^® CaM KII pAb was used in conjunction with an immunoperoxidase detection kit for immunohistochemical analysis.  For Western blot analysis, a 1:5000 dilution of the antibody was used.  (3202)

Notes: CaMKII_(281-309) was added to metaphase-arrested extracts. After adding calcium, the extracts were incubated at room temperature. Anti-ACTIVE^® CaMKII pAb and Anti-ACTIVE^® qualified HRP secondary antibodies were used to probe immunoblots for phospho-T²⁸⁶ CaMKIIα. (2590)

Notes: This paper studied the effects of both acute stress and Corticotropin-releasing factor (CRF) on hippocampus-dependent learning and on long-term synaptic plasticity in the mouse hippocampus.  In particular, the roles of PKC and CaMKII in the regulation of hippocampal long-term synaptic plasticity and in the performance of mice in a hippocampus-dependent learning task were studied. The level of active CaM KII in mouse hippocampal extracts was investigated by western blotting using the Anti-ACTIVE™ CaM KII antibody.  (2765)

Notes: The authors show how certain common tissue handling techniques, such as equilibration for 1-2 hours in a physiological medium and freezing, can dramatically affect the level of autonomous CaM KII activity in hippocampal tisuse, leading to potential artefacts in data. Activation of Cam KII was monitored by Western blot analysis using the Anti-ACTIVE® CaM KII pAb primary antibody. (2408)

Notes: The signal transduction mechanisms of adrenoreceptors in regulating K+ currents in isolated canine ventricular myocytes and human embryonic kidney (HEK 293) cells were examined. The CamKII pathway was monitored in the presence or absence of a specific inhibitor CaM KII inhibitor by Western blot using the Anti-ACTIVE® CaM KII pAb. (2407)

Notes: To demonstrate an interaction of activated CaM KII with the postsynaptic density protein densin-180, a gst-fusion protein of densin was produced and reacted with autophosphorylated CaM KII. The glutathione matrix-bound material was eluted and immunoblotted for phospho-Thr286-CaM KII. The autophosphorylated CaM KII was pulled down on the Glutathione only when the densin-GST fusion was present. The phosphoCaM KII was detected with the Anti-ACTIVE^® CaM KII pAb. The clone of densin was generated by RT-PCR of rat forebrain total RNA with the Access RT-PCR System. (0076)

Notes: Xenopus embryos were split into dorsal and ventral halves at stage 7. The total CaM KII was immunoprecipitated then blotted with the Anti-ACTIVE^® CaM KII pAb. Both halves had activated CaM KII but the ventral portion displayed a greater increase in phosphorylation. Stripping and reprobing the membranes with a normal CaM KII antibody demonstrated that equal amounts of CaM KII were present in the two samples. (0070)

Notes: Neurogranin deficient mice did not exhibit developmental or neuroanatomical abnormalities but had deficits in learning and memory. The levels of phosphorylated CaM KII in hippocampal extracts was determined by quantitative immunoblotting using the Anti-ACTIVE® CamKII pAb. (2410)

Notes: Treatment of  brain-derived neurotrophic factor increased MAPK-dependent synapsin I phosphorylation in rat cerebral cortex synaptosomal preparations. MAPK activity was determined by Western blot analysis using the Anti-ACTIVE® MAPK (1:10,000 dilution) pAb to detect the dually phosphorylated forms of MAPK. CaM kinase II activity was assayed by Western blot analysis with the Anti-ACTIVE® CaM KII pAb, Rabbit, (pT286) (1:2500 dilution). The Anti-Mouse IgG (H+L), AP Conjugate (1:1000 dilution) was used as a secondary antibody in Western blot analysis to detect TrkB expression. (2393)

Notes: The presence of autophosphorylated CaM KII in control or angiotensin II-treated neuronal cultures made from Sprague-Dawley rats was determined by Western blotting with a 1:5000 dilution of the Anti-ACTIVE^® CaMKII pAb. Cell Extracts were made with a buffer containing 1% NP-40, 10% glycerol, 150 mM NaCl, 20 mM Tris-HCl and protease inhibitors. (0063)

Notes: The Anti-ACTIVE^® CaM KII pAb was used to detect phospho-T²⁸⁶ of rat calmodulin kinase II in COS-7 cells by immunocytochemistry. Good detail is provided for the immunocytochemistry protocol. (1359)