Human STAT1 Baculovirus-Insect cells Overexpression Lysate


Human STAT1 Baculovirus-Insect cells Overexpression Lysate: 产品信息

This Human STAT1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of STAT1 protein (Cat: 12766-H20B) from the overexpression lysate was verified.
Baculovirus-Insect cells
A DNA sequence encoding the human STAT1 isoform beta (P42224-2) (Met 1-Val 712) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The recombinant human STAT1/GST chimera consists of 950 amino acids and has a calculated molecular mass of 111 kDa. It migrates as an approximately 105 kDa band in SDS-PAGE under reducing conditions.

Human STAT1 Baculovirus-Insect cells 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 STAT1 Baculovirus-Insect cells Overexpression Lysate: 别称

Human CANDF7 Overexpression Lysate; Human IMD31A Overexpression Lysate; Human IMD31B Overexpression Lysate; Human IMD31C Overexpression Lysate; Human ISGF-3 Overexpression Lysate; Human STAT91 Overexpression Lysate

STAT1 背景信息

STAT1 is a member of the STAT protein family. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor-associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. STAT1 can be activated by various ligands, including interferon-alpha, interferon-gamma, EGF, PDGF and IL6. It is a signal transducer and transcription activator that mediates cellular responses to interferons (IFNs), cytokine KITLG/SCF and other cytokines and growth factors. The phosphorylated STATs dimerize, associate with ISGF3G/IRF-9 to form a complex termed ISGF3 transcription factor, that enters the nucleus. ISGF3 binds to the IFN stimulated response element (ISRE) to activate the transcription of interferon-stimulated genes, which drive the cell in an antiviral state. In response to type II IFN (IFN-gamma), STAT1 is tyrosine- and serine-phosphorylated. It then forms a homodimer termed IFN-gamma-activated factor (GAF), migrates into the nucleus and binds to the IFN gamma activated sequence (GAS) to drive the expression of the target genes, inducing a cellular antiviral state. STAT1 becomes activated in response to KITLG/SCF and KIT signaling and may mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Defects in STAT1 can cause STAT1 deficiency complete and familial candidiasis type 7.
signal transducer and activator of transcription 1, 91kDa
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  • Shah S, et al. (2011) A novel disrupter of telomere silencing 1-like (DOT1L) interaction is required for signal transducer and activator of transcription 1 (STAT1)-activated gene expression. J Biol Chem. 286(48):41195-204.
  • Barthson J, et al. (2011) Cytokines tumor necrosis factor- and interferon-gamma induce pancreatic beta-cell apoptosis through STAT1-mediated Bim protein activation. J Biol Chem. 286(45):39632-43.
  • Xin Z, et al. (2011) PCBP2 enhances the antiviral activity of IFN- against HCV by stabilizing the mRNA of STAT1 and STAT2. PLoS One. 6(10):e25419.
  • Perwitasari O, et al. (2011) Inhibitor of B kinase epsilon (IKK(epsilon)), STAT1, and IFIT2 proteins define novel innate immune effector pathway against West Nile virus infection. J Biol Chem. 286(52):44412-23.
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