Human Protein Kinase D2/PRKD2 Baculovirus-Insect cells Overexpression Lysate: Product Information
This Human Protein Kinase D2/PRKD2 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Protein Kinase D2/PRKD2 protein (Cat: 11671-H20B) from the overexpression lysate was verified.
A DNA sequence encoding the amino acid sequence (Met 1-Leu 878) of human PRKD2 (NP_057541.2) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The recombinant human PRKD2/GST chimera consists of 1115 amino acids and has a calculated molecular mass of 124 kDa. It migrates as an approximately 120 kDa band in SDS-PAGE under reducing conditions.
Human Protein Kinase D2/PRKD2 Baculovirus-Insect cells Overexpression Lysate: Usage Guide
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.
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.
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human Protein Kinase D2/PRKD2 Baculovirus-Insect cells Overexpression Lysate: Alternative Names
Human HSPC187 Overexpression Lysate; Human nPKC-D2 Overexpression Lysate; Human PKD2 Overexpression Lysate
Protein Kinase D2/PRKD2 Background Information
Serine/threonine-protein kinase D2, also known as PRKD2 and PKD2, is a cytoplasm and membrane protein which belongs to theprotein kinase superfamily, CAMK Ser/Thr protein kinase family and PKD subfamily. PRKD2 / PKD2 is widely expressed. It contains onePH domain, twophorbol-ester/DAG-type zinc fingers and oneprotein kinase domain. PRKD2 / PKD2 is activated by DAG and phorbol esters. Phorbol-ester/DAG-type domains bind DAG, mediating translocation to membranes. Autophosphorylation of Ser-71 and phosphorylation of Ser-76 by PKC relieves auto-inhibition by the PH domain. PRKD2 / PKD2 converts transient diacylglycerol (DAG) signals into prolonged physiological effects, downstream of PKC. Involved in resistance to oxidative stress.
protein kinase D2
Zhang QH, et al., 2000, Genome Res 10 (10): 1546-60.
Di Bernardo MC, et al.,2008, Nature genetics. 40 (10) :1204-10.
Zhang L, et al., 2009, American journal of nephrology. 30 (6) :505-13.
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