This Human OX40 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of OX40 protein (Cat: 10481-H03H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Ala 216) of human TNFRSF4 (NP_003318.1) precursor was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
The recombinant human TNFRSF4/Fc is a disulfide-linked homodimer after removal of the signal peptide. The reduced monomer consists of 436 amino acids and has a predicted molecular mass of 48.2 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh TNFRSF4/Fc monomer is approximately 68 kDa due to glycosylation.
Human OX40 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 OX40 HEK293 Overexpression Lysate: 别称
Human ACT35 Overexpression Lysate; Human CD134 Overexpression Lysate; Human IMD16 Overexpression Lysate; Human OX40 Overexpression Lysate; Human TXGP1L Overexpression Lysate
OX4 (CD134) and its binding partner, OX4L (CD252), are members of the tumor necrosis factor receptor/tumor necrosis factor superfamily, is known to break an existing state of tolerance in malignancies, leading to a reactivation of antitumor immunity. The interaction between OX4 and OX4L plays an important role in antigen-specific T-cell expansion and survival. OX4 and OX4L also regulate cytokine production from T cells, antigen-presenting cells, natural killer cells, and natural killer T cells, and modulate cytokine receptor signaling. In line with these important modulatory functions, OX4-OX4L interactions have been found to play a central role in the development of multiple inflammatory and autoimmune diseases, making them attractive candidates for intervention in the clinic. Conversely, stimulating OX4 has shown it to be a candidate for therapeutic immunization strategies for cancer and infectious disease.
Compaan D.M., et al. (2006) .The crystal structure of the costimulatory OX40-OX40L complex. Structure 14:1321-1330.
Kawamata S., et al. (1998) .Activation of OX40 signal transduction pathways leads to tumor necrosis factor receptor-associated factor (TRAF) 2- and TRAF5-mediated NF-kappaB activation. J. Biol. Chem. 273:5808-5814.
Byun M., (2013) Inherited human OX40 deficiency underlying classic Kaposi sarcoma of childhood. J. Exp. Med. 210:1743-1759.