This Mouse CAMKIV overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of CAMKIV protein (Cat: 50838-M20B) from the overexpression lysate was verified.
A DNA sequence encoding the mouse CAMK4 (P08414) (Met1-Tyr469) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The secreted recombinant mouse CAMK4/GST chimera consists of 706 amino acids and has a calculated molecular mass of 80.4 kDa. The recombinant protein migrates as an approximately 85 kDa band in SDS-PAGE under reducing conditions.
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Ca2+/ calmodulin-dependent protein kinase 4 (CAMKⅣ) belongs to the serine/threonine protein kinase family, and to the Ca2+/calmodulin-dependent protein kinase subfamily which is widely recognized as an essential enzyme implicated in the phophoinositide amplification cascade. Ca2+/calmodulin dependent protein kinase (CAMK) can be activated by the introcellular increased Ca2+ and then apt to combine with the target protein. Ca2+/ calmodulin-dependent protein kinase 4 (CAMKⅣ) is a multifunctional CaM-dependent kinase protein with limited tissue distribution, that has been implicated in transcriptional regulation in lymphocytes, neurons and male germ cells. All of the isforms of this family, including myosin light chain kinase, phosphorylase kinase, CaMK1, CaMKⅢ and CaMKⅣ have EF-hand structure.
calcium/calmodulin-dependent protein kinase IV
EGFR Signaling Pathway
B Cell Receptor Signaling Pathway
G Protein-coupled Receptors Signaling
T Cell Receptor Signaling Pathway
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