Human NAALADL1 HEK293 Overexpression Lysate

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Human NAALADL1 HEK293 Overexpression Lysate: Product Information

Product Description
This Human NAALADL1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NAALADL1 protein (Cat: 11146-H07H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the extracellular domain of human NAALADL1 (NP_005459.2) (Pro 29-Leu 740) was expressed, with a polyhistidine tag at the N-terminus.
Molecule Mass
The recombinant human NAALADL1comprises 728 amino acids and has a predicted molecular mass of 80 kDa. As a result of glycosylation, rh NAALADL1 migrates as an approximately 90 kDa band in SDS-PAGE under reducing conditions.

Human NAALADL1 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 NAALADL1 HEK293 Overexpression Lysate: Alternative Names

Human I100 Overexpression Lysate; Human NAALADASEL Overexpression Lysate; Human NAALADL1 Overexpression Lysate

NAALADL1 Background Information

N-acetylated-alpha-linked acidic dipeptidase-like protein, also known as NAALADL1, NAALADase L, and Ileal dipeptidylpeptidase, is a Single-pass type I I membrane protein and a member of the peptidase M28 family and M28B subfamily. NAALADase L is mainly expressed in the distal small intestine. It is also expressed in the spleen and testis and Weakly expressed in the brain, locating mainly to the brain stem, amygdala, thalamus and ventral striatum. NAALADase L is a chloride-activated, membrane bound, metallopeptidase that cleaves the endogenous neuropeptide N-acetyl-aspartyl-glutamate (NAAG). NAAG acts as a partial NMDA agonist as well as a full agonist at the presynaptic metabotropic glutamate receptor 3 (mGluR3), where it acts to reduce glutamate release. NAALADase L also exhibits a dipeptidyl-peptidase IV type activity. NAALADase inhibition may be a novel therapeutic approach to reduce or inhibit heightened aggressiveness, and possibly to treat aggressive behavior associated with psychiatric disorders.
Full Name
N-acetylated alpha-linked acidic dipeptidase-like 1
References
  • Stauch BL. 1989, Neurosci Lett. 100 (1-3): 295-300.
  • Shneider BL. et al., 1997, J. Biol. Chem. 272: 31006-15.
  • Pangalos MN. et al., 1999, J. Biol. Chem. 274: 8470-83.
  • Thomas AG. et al., 1999, Brain Res. 843 (1-2): 48-52.
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