Human IL4R HEK293 Overexpression Lysate: Product Information
This Human IL4R overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IL4R protein (Cat: 10402-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human IL4R (NP_000409.1) precursor (Met 1-His 232) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant human IL4R consists of 218 amino acids after cleavage of the signal peptide and predictes a molecular mass of 25.3 kDa. In SDS-PAGE under reducing conditions, it migrates with the apparent molecular mass of 43-48 kDa due to glycosylation.
Human IL4R HEK293 Overexpression Lysate: Usage Guide
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).
Stability & Storage
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 IL4R HEK293 Overexpression Lysate: Alternative Names
Human CD124 Overexpression Lysate; Human IL-4RA Overexpression Lysate; Human IL4R Overexpression Lysate; Human IL4RA Overexpression Lysate
IL4R Background Information
The cluster of differentiation (CD) system is commonly used as cell markers in immunophynotyping. Different kinds of cells in the immune system can be identified through the surface CD molecules which associating with the immune function of the cell. There are more than 32 CD unique clusters and subclusters have been identified. Some of the CD molecules serve as receptors or ligands important to the cell through initiating a signal cascade which then alter the behavior of the cell. Some CD proteins do not take part in cell signal process but have other functions such as cell adhesion. CD124, also known as interleukin 4 receptor (IL4R), is a typeⅠ transmembrane protein that can regulate IgE antibody production in B cells through binding to interleukin 4 and interleukin 13 and promote differentiation of Th2 cells through binding to interleukin 4. The membrane-bound form of CD124 can be hydrolyzed to soluble form which can inhibit IL4-mediated cell proliferation and IL5 upregulation by T-cells.
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