Human METAP1 HEK293 Overexpression Lysate

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Human METAP1 HEK293 Overexpression Lysate: Product Information

Product Description
This Human METAP1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of METAP1 protein (Cat: 10241-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the N-terminal domain (Met 1-Phe 272) of human METAP1 (NP_055958.2) was fused with the Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human Fc/METAP1 is a disulfide-linked homodimeric protein. The reduced monomer consists of 509 amino acids and has a predicted molecular mass of 57 kDa. As a result of glycosylation, the apparent molecular mass of rh Fc/METAP1 monomer is approximately 60-65 kDa in SDS-PAGE under reducing conditions.

Human METAP1 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
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.
Recommend Usage
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.
Sample Buffer
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human METAP1 HEK293 Overexpression Lysate: Alternative Names

Human MAP1A Overexpression Lysate; Human MetAP1A Overexpression Lysate

METAP1 Background Information

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.
Full Name
methionyl aminopeptidase 1
References

    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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