This Human GBP1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of GBP1 protein (Cat: 12569-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human GBP1 (AAA35871.1) (Met 1-Cys 589) was fused with a polyhistidine tag at the C-terminus.
The recombinant human GBP1 comprises 600 amino acids and has a predicted molecular mass of 69 kDa. rhGBP1 migrates as an approximately 65 kDa band in SDS-PAGE under reducing conditions.
Human GBP1 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.
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 GBP1 HEK293 Overexpression Lysate: 别称
Human GBP1 Overexpression Lysate
Guanylate-binding protein 1 (GBP-1) is a member of the GBP family whose members are GTPases induced in response to interferon-λ (IFN-λ), with seven highly homologous members in humans, termed HuGBP-1 to HuGBP-7. GBP-1 expression is induced by type1 and type2 interferons, including IFN-λ and also by interleukin-1β (IL-1β), IL-1α, and tumor necrosis factor-α (TNF-α). GBP-1 is key to the protective immunity against microbial and viral pathogens. GBP-1 was only secreted from endothelial cells. Secretion occurred without the presence of a leader peptide. Secretion procession is a nonclassical, likely ABC transporter-dependent, pathway and independent of GBP-1 GTPase activity and isoprenylation, and did not require additional interferon-λ-induced factors. Clinically most important was the detection of significantly increased GBP-1 concentrations in the cerebrospinal fluid of patients with bacterial meningitis as compared to control patients.
guanylate binding protein 1
NF-kB (NFkB) Pathway
Tripal P, et al. (2007) Unique features of different members of the human guanylate-binding protein family. J Interferon Cytokine Res. 27(1): 44-52.
Naschberger E, et al. (2006) Human Guanylate Binding Protein-1 Is a Secreted GTPase Present in Increased Concentrations in the Cerebrospinal Fluid of Patients with Bacterial Meningitis. Am J Pathol. 169(3): 1088-99.