Human PRKAR1A HEK293 Overexpression Lysate

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Human PRKAR1A HEK293 Overexpression Lysate: 产品信息

产品描述
This Human PRKAR1A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PRKAR1A protein (Cat: 14416-H08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human PRKAR1A (P10644) (Met1-Val381) was expressed with a polyhistidine tag at the C-terminus.
分子量
The recombinant human PRKAR1A consists of 391 amino acids and predicts a molecular mass of 44.3 KDa. It migrates as an approximately 47 KDa band in SDS-PAGE under reducing conditions.

Human PRKAR1A HEK293 Overexpression Lysate: 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human PRKAR1A HEK293 Overexpression Lysate: 别称

Human ACRDYS1 Overexpression Lysate; Human ADOHR Overexpression Lysate; Human CAR Overexpression Lysate; Human CNC Overexpression Lysate; Human CNC1 Overexpression Lysate; Human PKR1 Overexpression Lysate; Human PPNAD1 Overexpression Lysate; Human PRKAR1 Overexpression Lysate; Human TSE1 Overexpression Lysate

PRKAR1A 背景信息

PRKAR1A, also known as PRKAR1 and PKR1, is one of the regulatory subunits of cAMP-dependent protein kinase A (PKA). PKA can be activated by cAMP. cAMP is a signaling molecule important for a variety of cellular functions. cAMP exerts its effects by activating PKA, which transduces the signal throughphosphorylation of different target proteins. The inactive holoenzyme of PKA is a tetramer composed of two regulatory and two catalytic subunits. cAMP causes the dissociation of the inactive holoenzyme into a dimer of regulatory subunits bound to four cAMP and two free monomeric catalytic subunits. Four different regulatory subunits and three catalytic subunits of PKA have been identified in humans. PRKAR1A was found to be a tissue-specific extinguisher that down-regulates the expression of seven liver genes in hepatoma x fibroblast hybrids Three alternatively spliced transcript variants encoding the same protein have been observed.
全称
protein kinase, cAMP-dependent, regulatory, type I, alpha
参考文献
  • Huang L J. et al., 1997, Proc Natl Acad Sci. 94 (21): 11184-9.
  • Herberg F W. et al., 2000, J Mol Biol. 298 (2): 329-39.
  • Scambler P. et al., 1987, Am J Hum Genet. 41 (5): 925-32.
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