Human methionyl aminopeptidase 2/METAP2 Baculovirus-Insect Overexpression Lysate

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Human methionyl aminopeptidase 2/METAP2 Baculovirus-Insect Overexpression Lysate: 製品の情報

製品の説明
This Human methionyl aminopeptidase 2/METAP2 overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of methionyl aminopeptidase 2/METAP2 protein (Cat: 10245-H08B) from the overexpression lysate was verified.
発現ホスト
Baculovirus-Insect Cells
Human
タンパク質構築情報
A DNA sequence encoding the human METAP2 (NP_006829.1) (Met1-Tyr478) was expressed with a polyhistidine tag at the C-terminus.
分子量
The recombinant human METAP2 consists 489 amino acids and predicts a molecular mass of 54.3 kDa.

Human methionyl aminopeptidase 2/METAP2 Baculovirus-Insect Overexpression Lysate: 用法

調製方法
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.
溶解バッファー
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.
おすすめの用法
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.
バッファー
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
安定性 & 保存条件
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.
アプリケーション
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human methionyl aminopeptidase 2/METAP2 Baculovirus-Insect Overexpression Lysate: 別名

Human MAP2 Overexpression Lysate; Human MNPEP Overexpression Lysate; Human p67 Overexpression Lysate; Human p67eIF2 Overexpression Lysate

methionyl aminopeptidase 2/METAP2 背景情報

METAP2 (Methionine aminopeptidase 2), also known as MAP2 is a a protein which belongs to the peptidase M24A family. MAP2 binds 2 cobalt or manganese ions and contains approximately 12 O-linked N-acetylglucosamine (GlcNAc) residues. It is found in all organisms and is especially important because of its critical role in tissue repair and protein degradation. The catalytic activity of human MAP2 toward Met-Val peptides is consistently two orders of magnitude higher than that of METAP1, suggesting that it is responsible for processing proteins containing N-terminal Met-Val and Met-Thr sequences in vivo. This protein functions both by protecting the alpha subunit of eukaryotic initiation factor 2 from inhibitory phosphorylation and by removing the amino-terminal methionine residue from nascent protein. MAP2 protects eukaryotic initiation factor EIF2S1 from translation-inhibiting phosphorylation by inhibitory kinases such as EIF2AK2/PKR and EIF2AK1/HCR. It also plays a critical role in the regulation of protein synthesis.
完全な名称
methionyl aminopeptidase 2
参考文献
  • Bennett, et al. (1997) EPR Studies on the Mono- and Dicobalt (II)-Substituted Forms of the Aminopeptidase from Aeromonas proteolytica. Insight into the Catalytic Mechanism of Dinuclear Hydrolases. J Am Chem Soc. 119:1923-33.
  • Johansson, et al. (2008) Dicobalt II-II, II-III, and III-III Complexes as Spectroscopic Models for Dicobalt Enzyme Active Sites. Inorg Chem. 47:5079-92.
  • Bradshaw, et al. (2002) Aminopeptidases and angiogenesis. Essays Biochem. 38: 5-78.
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