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Synthetic phosphopeptide derived from a region of human CaM Kinase II alpha that contains threonine 286.
Calcium/calmodulin-dependent protein kinase II alpha (CaM Kinase II alpha) is a 50 kDa member of CaM Kinase II family of serine-threonine kinases that transduce Ca2+ signals to several target proteins, including ion channels and transcription activators. CaM Kinase II is predominantly expressed in two isoforms in the brain: alpha and beta. CaM Kinase II plays an important role in neuronal plasticity and memory formation, and exerts both calcium-calmodulin-dependent and -independent activities. Autophosphorylation of CaM Kinase II alpha on threonine 286 allows the kinase to switch from a calmodulin-dependent to a calmodulin-independent state, and is required for various cellular functions including hippocampal long-term potentiation (LTP), special learning, and hippocampus-dependent memory.
Our Abpromise guarantee covers the use of ab5683 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||1/1000. Detects a band of approximately 50 kDa.|
|IHC (PFA fixed)||1/300.|
Immunofluorescent staining for CaMKII alpha phospho (T286) using ab5683 (1/300, incubated for 18 hours) in rat spinal cord. To induce CaMKII alpha phospho (T286) protein expression, a noxious stimulus was administered to the rat 5 minutes prior to 4% PFA perfusion fixation (this is a known paradigm for inducing phosphorylation CaMKII in some spinal neurons). The resulting immunofluorescent staining for CaMKII alpha phospho (T286) protein is observed in the cytoplasm of many dorsal horn spinal neurons (surrounding the central canal [A] or in the ventral horn [B]). Omition of primary antibody resulted in a lack of staining (data not shown).
Protocol details: Tissue was prepared by 4% paraformaldehyde cardiac perfusion fixation. Tissue was frozen on dry ice and then embedded in OCT compound and cut on cryostat. An antigen retrieval step was not neccesary for the IHC protocol.
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