Human PRAK/MAPKAPK5 Baculovirus-Insect cells Overexpression Lysate: 产品信息
This Human PRAK/MAPKAPK5 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of PRAK/MAPKAPK5 protein (Cat: 13655-H20B) from the overexpression lysate was verified.
A DNA sequence encoding the human MAPKAPK5 isoform 1 (NP_003659.2) (Met 1-Gln 471) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The recombinant human MAPKAPK5/GST chimera consists of 708 amino acids and has a calculated molecular mass of 82 KDa. It migrates as an approximately 75KDa band in SDS-PAGE under reducing conditions.
Human PRAK/MAPKAPK5 Baculovirus-Insect cells Overexpression Lysate: 使用指南
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).
稳定性 & 储存条件
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 PRAK/MAPKAPK5 Baculovirus-Insect cells Overexpression Lysate: 别称
Human MAPKAP-K5 Overexpression Lysate; Human MK-5 Overexpression Lysate; Human MK5 Overexpression Lysate; Human PRAK Overexpression Lysate
MAPKAPK5 contains 1 protein kinase domain and belongs to the protein kinase superfamily, CAMK Ser/Thr protein kinase family. MAPKAPK5 has significant sequence homology to mitogen-activated protein kinase (MAPK)-activated protein kinase (MAPKAPK). It is widely distributed. MAPKAPK5 can be phosphorylated by an extracellular-regulated kinase (ERK), and p38 kinase but not by c-jun N-terminal kinase (JNK)in vitro. Recombinant GST-MAPKAPK5 protein can phosphorylate a peptide derived from the regulatory light chain of myosin II. Phosphorylation of MAPKAPK5 by ERK and p38 kinase increased its activity by 9 and 15 fold respectively. Taken together, these data suggest that MAPKAPK5 is a novel in vitro substrate for ERK and p38 kinase. In response to cellular stress and proinflammatory cytokines, this kinase is activated through its phosphorylation by MAP kinases including MAPK1/ERK, MAPK14/p38-alpha, and MAPK11/p38-beta. MAPKAPK5 also mediates stress-induced small heat shock protein 27 phosphorylation.
mitogen-activated protein kinase-activated protein kinase 5
P38 MAPK Signaling Pathway
Ni H, et al. (1998) MAPKAPK5, a novel mitogen-activated protein kinase (MAPK)-activated protein kinase, is a substrate of the extracellular-regulated kinase (ERK) and p38 kinase. Biochem Biophys. Res Commun. 243 (2): 492-6.
Sudo T, et al. (2000) p62 functions as a p38 MAP kinase regulator. Biochem Biophys Res Commun. 269 (2): 521-5.
Strausberg RL, et al. (2003) Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. Proc Natl Acad Sci. 99 (26): 16899-903.
New L, et al. (2004) Regulation of PRAK subcellular location by p38 MAP kinases. Mol Biol Cell. 14 (6): 2603-16.