Human CaMKII beta/CAMK2B Baculovirus-Insect cells Overexpression Lysate

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Human CaMKII beta/CAMK2B Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human CaMKII beta/CAMK2B overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of CaMKII beta/CAMK2B protein (Cat: 11738-H20B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human CAMK2B isoform 2 (Q13554-3) (Met 1-Gln 503) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Molecule Mass
The recombinant human CAMK2B/GST chimera consists of 740 amino acids and has a calculated molecular mass of 84.2 kDa. It migrates as an approximately 85 kDa band in SDS-PAGE under reducing conditions.

Human CaMKII beta/CAMK2B Baculovirus-Insect cells Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human CaMKII beta/CAMK2B Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human CAM2 Overexpression Lysate; Human CAMK2 Overexpression Lysate; Human CAMKB Overexpression Lysate

CaMKII beta/CAMK2B Background Information

Calcium/calmodulin-dependent protein kinase II beta (CAMK2B) is a member of the serine/threonine protein kinase family and to the Ca(2+)/calmodulin-dependent protein kinase subfamily. CaMKII is an important player in prostate cancer cells ability to escape apoptosis under androgen ablation and facilitate the progression of prostate cancer cells to an androgen independent state. As a multifunctional protein kinase, the loss of activity may play a critical role in initiating the changes leading to ischemia-induced cell death. CaMKII are found to be important for the functions of immune cells. CaMKII can be activated by TLR ligands, and in turn promotes both myeloid differentiating factor 88 and Toll/IL-1 receptor domain-containing adaptor protein-inducing IFN-beta-dependent inflammatory responses by directly activating TAK1 and IRF3. CAMKII has four subunit isoforms (alpha, beta, gamma, delta). It is possible that distinct isoforms of this chain have different cellular localizations and interact differently with calmodulin. The alpha- and beta-isoforms have narrow distributions restricted mainly to neuronal tissues, but the gamma- and delta-isoforms are ubiquitously expressed within neuronal and non-neuronal tissues. CAMK2B is important for controlling the direction of plasticity at the parallel fiber-Purkinje cell synapse. CaMK2 is involved in neuronal survival through the reorganization of the neuroarchitecture and that the regulation of this role is controlled at the level of gene expression. Because CaMK2B influences the expression of many neuroreceptors and influences neural outgrowth and pruning, its altered expression in the cerebral cortex in schizophrenia or depression may contribute to schizophrenia and depression.
Full Name
calcium/calmodulin-dependent protein kinase II beta
References
  • Hiestand DM, et al. (1992)Calcium/calmodulin dependent protein kinase II mRNA in the gerbil brain after cerebral ischemia. Neurosci Lett. 144(1-2): 75-8.
  • Shackelford DA, et al. (1995) Effect of cerebral ischemia on calcium/calmodulin-dependent protein kinase II activity and phosphorylation. J Cereb Blood Flow Metab. 15(3): 450-61.
  • Walikonis R S, et al. (2001) Densin-180 forms a ternary complex with the (alpha)-subunit of Ca2+/calmodulin-dependent protein kinase II and (alpha)-actinin. J. Neurosci. (United States). 21 (2): 423-33.
  • Shimazaki A, et al. (2006) Calcium/calmodulin-dependent protein kinase II in human articular chondrocytes. Biorheology. 43(3-4): 223-33.
  • Novak G, et al. (2006) Increased expression of calcium/calmodulin-dependent protein kinase IIbeta in frontal cortex in schizophrenia and depression. Synapse. 59(1): 61-8.
  • Rokhlin OW, et al. (2007) Calcium/calmodulin-dependent kinase II plays an important role in prostate cancer cell survival. Cancer Biol Ther. 6(5): 732-42.
  • van Woerden GM, et al. (2009) betaCaMKII controls the direction of plasticity at parallel fiber-Purkinje cell synapses. Nat Neurosci. 2009 Jul;12(7): 823-5. .
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