Neosartorya fumigata MITF HEK293 Overexpression Lysate

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Neosartorya fumigata MITF HEK293 Overexpression Lysate: 产品信息

产品描述
This Neosartorya fumigata MITF overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of MITF protein (Cat: 69009-A08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Neosartorya fumigata
蛋白构建信息
A DNA sequence encoding the mitF (CAA41217.1) (Met1-His176) was expressed with a polyhistidine tag at the C-terminus.
分子量
The recombinant mitF consists of 160 amino acids and predicts a molecular mass of 18.3 kDa.

Neosartorya fumigata MITF HEK293 Overexpression Lysate: 使用指南

制备方法
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.
裂解缓冲液
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.
使用建议
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.
缓冲液
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.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

MITF 背景信息

Microphthalmia-associated transcription factor (MITF) is a member of the basic helix-loop-helix leucine zipper (bHLH-Zip) family and functions as the master regulator of the melanocytic lineage.MITF (Microphthalmia-associated transcription factor) is a lineage-specific transcription factor that plays a critical role in melanocyte homeostasis and whose deregulation has been shown to contribute to melanoma disease. Microphthalmia-associated transcription factor (MITF) is expressed in melanomas and has a critical role in melanocyte development and transformation. Because inhibition of MITF inhibits cell growth in melanoma, MITF is a potential therapeutic target molecule. Microphthalmia-associated transcription factor (MITF) regulates the transcription of its target genes by binding to their promoters. Microphthalmia-associated transcription factor (MITF) is a key regulator of differentiation of melanocytes and retinal pigment epithelial cells, but it also has functions in non-pigment cells.
全称
microphthalmia-associated transcription factor
Related Pathways
  • MAPK-Erk Pathway
    MAPK-Erk Pathway
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