Human VTI1A HEK293 Overexpression Lysate


Human VTI1A HEK293 Overexpression Lysate: 产品信息

This Human VTI1A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of VTI1A protein (Cat: 14595-H04H) from the overexpression lysate was verified.
HEK293 Cells
A DNA sequence encoding the human VTI1A (NP_660207.2) (Met1-Arg187) was expressed with the Fc region of mouse IgG1 at the N-terminus.
The recombinant human VTI1A consists 423 amino acids and predicts a molecular mass of 48.5 kDa.

Human VTI1A HEK293 Overexpression Lysate: 使用指南

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.
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
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.
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human VTI1A HEK293 Overexpression Lysate: 别称

Human MMDS3 Overexpression Lysate; Human MVti1 Overexpression Lysate; Human Vti1-rp2 Overexpression Lysate; Human VTI1RP2 Overexpression Lysate

VTI1A 背景信息

Evolutionarily conserved SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptors) proteins form a complex that drives membrane fusion in eukaryotes. VTI1A and VTI1B are the only members of their SNARE subclass and the yeast homolog Vti1p is essential for cell survival. VTI1A is involved in regulating insulin-stimulated trafficking of secretory vesicles enriched with both GLUT4 (glucose transporter) and Acrp30 in adipocytes; it also plays key roles in neuronal development and in selectively maintaining spontaneous neurotransmitter release.
vesicle transport through interaction with t-SNAREs 1A
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