Human LSAMP HEK293 Overexpression Lysate

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Human LSAMP HEK293 Overexpression Lysate: Product Information

Product Description
This Human LSAMP overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of LSAMP protein (Cat: 12136-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human LSAMP (Q13449) (Met 1-Asn 315), without the pro peptide, was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The secreted mature form of recombinant human LSAMP consists of 298 amino acids and has a predicted molecular mass of 33.4 kDa. As a result of glycosylation, the apparent molecular mass of rhLSAMP is approximately 45-55 kDa in SDS-PAGE under reducing conditions.

Human LSAMP 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 LSAMP HEK293 Overexpression Lysate: Alternative Names

Human FLJ34254 Overexpression Lysate; Human FLJ35396 Overexpression Lysate; Human FLJ37216 Overexpression Lysate; Human FLJ54658 Overexpression Lysate; Human IGLON3 Overexpression Lysate; Human LAMP Overexpression Lysate

LSAMP Background Information

The limbic system-associated membrane protein (LAMP) is a cell surface glycoprotein expressed by cortical and subcortical regions of the mammalian CNS that comprise or receive direct projections from limbic system structures. The 64-68-kDa glycoprotein limbic system-associated membrane protein (LsAMP) is expressed on the surface of somata and proximal dendrites of neurons. These areas perform cognitive and autonomic functions, also learning and memory. The functional analysis indicates that LsAMP acts as a selective adhesion molecule, serving as a guidance cue for specific patterns of connectivity, which underlies the normal development of the limbic system. In animal studies there have been found that rats with increased level of anxiety had 1.6-fold higher expression of LsAMP gene in the periaqueductal gray compared to rats with low level of anxiety, indicating a possible role of LsAMP in the regulation of anxiety.
Full Name
limbic system-associated membrane protein
References
  • Zacco A. et al., 1990, The Journal of Neuroscience. 10(1): 73-90.
  • Pimenta AF. et al., 1996, Gene. 170(2) :189-95.
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