This Human CD27 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD27 protein (Cat: 10039-H03H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Ile 192) of human CD27 (NP_001233.1) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the carboxy-terminus.
The recombinant mature human CD27/Fc is a disulfide-linked homodimeric protein. The reduced monomer comprises 420 amino acids and has a calculated molecular mass of 47.2 kDa. As a result of glycosylation, the monomer migrates as an approximately 65 kDa band in SDS-PAGE under reducing conditions.
Human CD27 HEK293 Overexpression Lysate: 用法
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).
安定性 & 保存条件
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 CD27 HEK293 Overexpression Lysate: 別名
Human S152 Overexpression Lysate; Human S152. LPFS2 Overexpression Lysate; Human S152.LPFS2 Overexpression Lysate; Human T14 Overexpression Lysate; Human TNFRSF7 Overexpression Lysate; Human Tp55 Overexpression Lysate
CD27, also known as TNFRSF7, is a member of the TNF-receptor superfamily limited to cells of the lymphoid lineage, and exists as both a dimeric glycoprotein on the cell surface and as a soluble protein in serum. As a type I transmembrane glycoprotein of about 55 kDa existing as disulfide-linked homodimer, CD27 has been shown to play roles in lymphoid proliferation, differentiation, and apoptosis.It has an important role in the generation of T cell immunity and is an robust marker for normal memory B cells. It is a T and B cell co-stimulatory molecule, the activity of CD27 is governed by its TNF-like ligand CD70 on lymphocytes and dendritic cells. The CD27-CD70 interaction is required for Th1 generation responses to differentiation signals and long-term maintenance of T cell immunity, and meanwhile, plays a key role in regulating B-cell differentiation, activation and immunoglobulin synthesis.