Human Latexin HEK293 Overexpression Lysate 产品信息
Product Description
This Human Latexin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Latexin protein (Cat: 10211-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the human Latexin (NP_064554.3) (Glu 2-Glu 222) was fused with the Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human Fc/LXN is a disulfide-linked homodimeric protein.The reduced monomer consists of 458 amino acids and predicts a molecular mass of 52 kDa. As a result of glycosylation, the apparent molecular mass of rhFc/LXN monomer is approximately 60-65 kDa in SDS-PAGE under reducing conditions.
Human Latexin 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 Latexin HEK293 Overexpression Lysate Alternative Names
Human ECI Overexpression Lysate;Human TCI Overexpression Lysate
Latexin Background Information
Latexin, also known as endogenous carboxypeptidase inhibitor, tissue carboxypeptidase inhibitor, TCI, ECI and LXN, is a cytoplasm protein which belongs to the protease inhibitor I47 (latexin) family. It is highly expressed in heart, prostate, ovary, kidney, pancreas, and colon. Latexin / LXN is the only known endogenous specific inhibitor of zinc-dependent metallocarboxypeptidases (MCPs) present in mammalians so far. Latexin is originally identified as a molecular marker for the regional specification of the neocortex in development in rats. The 222 amino acid latexin in human shows different expression distribution with high levels in heart, prostate, ovary, kidney, pancreas, and colon, but only moderate or low levels in other tissues including brain. Latexin is also expressed at high levels and is inducible in macrophages in concert with other protease inhibitors and potential protease targets, and thus is suggested to play a role in inflammation and innate immunity pathways. Despite of the non-detectable sequence similarity with plant and parasite inhibitors, Latexin is related to a human putative tumor suppressor protein, TIG1. In addition, Latexin is also implicated in Alzheimer's disease.
References
Liu Q., et al.,(2000), Cloning, tissue expression pattern and genomic organization of latexin, a human homologue of rat carboxypeptidase A inhibitor. Mol. Biol. Rep. 27:241-246. Gauci S., et al., (2009), Lys-N and trypsin cover complementary parts of the phosphoproteome in a refined SCX-based approach.nal. Chem. 81:4493-4501. Muzny D.M., et al.,(2006), The DNA sequence, annotation and analysis of human chromosome 3.Nature 440:1194-1198.
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