Human HAI-2/SPINT2 HEK293 Overexpression Lysate

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Human HAI-2/SPINT2 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human HAI-2/SPINT2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of HAI-2/SPINT2 protein (Cat: 10324-H08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human SPINT2 (NP_066925.1) (Met1-Lys197) was expressed with a polyhistidine tag at the C-terminus.
分子量
The recombinant human SPINT2 consists 181 amino acids and predicts a molecular mass of 20.6 kDa.

Human HAI-2/SPINT2 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.

Human HAI-2/SPINT2 HEK293 Overexpression Lysate: 别称

Human DIAR3 Overexpression Lysate; Human HAI-2 Overexpression Lysate; Human HAI2 Overexpression Lysate; Human Kop Overexpression Lysate; Human PB Overexpression Lysate

HAI-2/SPINT2 背景信息

Choanal (CA) and gastrointestinal atresias (GA) are an important feature of syndromic congenital sodium diarrhea (sCSD), a disorder recently associated with mutations in the gene for serine protease inhibitor type 2 (SPINT2). The SPINT2 gene is epigenetically silenced or downregulated in human cancers, altering the balance of HGF activation/inhibition ratio, which contributes to cancer development and progression. SPINT2 is a tumor suppressor gene that inhibits proteases implicated in cancer progression, like HGFA, hepsin and matriptase. Loss of SPINT2 expression in tumors has been associated with gene promoter hypermethylation. SPINT2 (serine peptidase inhibitor Kunitz type 2), a proteolytic inhibitor of hepatocyte growth factor activator (HGFA), has a significant role in the suppression of the HGF-MET pathway and malignant melanoma progression.
全称
serine peptidase inhibitor, Kunitz type, 2
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