Human CD40 HEK293 Overexpression Lysate: Product Information
This Human CD40 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD40 protein (Cat: 10774-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human CD40 (NP_001241.1) extracellular domain (Met 1-Arg 193) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant human CD40 comprises 184 amino acids with a predicted molecular mass of 20.7 kDa. As a result of glycosylation, rhCD40 migrates as an approximately 32 kDa band in SDS-PAGE under reducing conditions.
Human CD40 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 CD40 HEK293 Overexpression Lysate: Alternative Names
Human Bp50 Overexpression Lysate; Human CDW40 Overexpression Lysate; Human p50 Overexpression Lysate; Human TNFRSF5 Overexpression Lysate
CD40 Background Information
CD40, also known as TNFRSF5, is a member of the TNF receptor superfamily which are single transmembrane-spanning glycoproteins. CD40 protein plays an essential role in mediating a broad variety of immune and inflammatory responses including T cell-dependent immunoglobulin class switching, memory B cell development, and germinal center formation. CD40 protein is expressed in B cells, dendritic cells, macrophages, endothelial cells, and several tumor cell lines. Defects in CD40 result in hyper-IgM immunodeficiency type 3 (HIGM3). In addition, CD40/CD40L interaction is found to be necessary for amyloid-beta-induced microglial activation, and thus is thought to be an early event in Alzheimer disease pathogenesis.
Successfully added to cart Please enter catalog numberSubmitted successfullyNetwork ErrorPlease enter your company namePlease enter your namePlease enter your emailPlease enter a valid email addressPlease enter some messageNot found.