Human METAP1 Baculovirus-Insect Overexpression Lysate

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Human METAP1 Baculovirus-Insect Overexpression Lysate: 产品信息

产品描述
This Human METAP1 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of METAP1 protein (Cat: 10241-H08B) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human METAP1 (NP_055958.2) (His52-Phe386) was expressed with a polyhistidine tag at the C-terminus and an initial Met at the N-terminus.
分子量
The recombinant human METAP1 consists 347 amino acids and predicts a molecular mass of 39.3 kDa.

Human METAP1 Baculovirus-Insect 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 METAP1 Baculovirus-Insect Overexpression Lysate: 别称

Human MAP1A Overexpression Lysate; Human MetAP1A Overexpression Lysate

METAP1 背景信息

Processing of the N-terminal initiator methionine or formylated methionine is an essential cellular process conserved from prokaryotes to eukaryotes. The proteolytic removal of N-terminal methionine from nascent peptides is catalyzed by a family of enzymes known as methionine aminopeptidases (MetAPs) and is essential for cell growth. METAP1 and METAP2 have different substrate specificity due to the differences in both size and shape of the active sites. As a member of the M24 family of metalloproteases, METAP1 plays an important role in G(2)/M phase regulation of the cell cycle and may serve as a promising target for the discovery and development of new anticancer agents.
全称
methionyl aminopeptidase 1
参考文献

    1. Lowther, W.T. and B.W. Matthews, 2000, Biochim. Biophys. Acta. 1477: 157 – 167.

    2. Addlaqatta, A. et al., 2005, Biochemistry. 44: 14741-14749.

    3. Hu, X. et al., 2006, Proc. Natl. Acad. Sci. U.S.A. 103:18148-18153.

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