Human CD30/TNFRSF8 HEK293 Overexpression Lysate: Product Information
This Human CD30/TNFRSF8 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD30/TNFRSF8 protein (Cat: 10777-H03H) from the overexpression lysate was verified.
A DNA sequence encoding the human TNFRSF8 (NP_001234.2) extracellular domain (Met 1-Lys 379) was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
The recombinant human TNFRSF8/Fc is a disulfide-linked homodimer after removal of the signal peptide. The reduced monomer consists of 609 amino acids and has a predicted molecular mass of 66.5 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhTNFRSF8/Fc monomer is approximately 130 kDa due to glycosylation.
Human CD30/TNFRSF8 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 CD30/TNFRSF8 HEK293 Overexpression Lysate: Alternative Names
Human CD30 Overexpression Lysate; Human D1S166E Overexpression Lysate; Human Ki-1 Overexpression Lysate
CD30/TNFRSF8 Background Information
CD3, also known as TNFRSF8, is a cell membrane protein of the tumor necrosis factor receptor (TNFR) superfamily. CD3 protein is expressed by activated, but not resting, T and B cells. CD3 can regulate proliferation of lymphocytes and may also play an important role in human immunodeficiency virus replication. As a regulator of apoptosis, CD3 protein induces cell death or proliferation, depending on the cell type, and has been shown to limit the proliferative potential of autoreactive CD8 effector T cells and protect the body against autoimmunity. CD3 protein expression is upregulated in various hematological malignancies, including Reed-Sternberg cells in Hodgkin's disease (HD), anaplastic large cell lymphoma (ALCL) and subsets of Non-Hodgkin's lymphomas (NHLs), and CD3 is also linked to leukocytes in patients with chronic inflammatory diseases, including lupus erythematosus, asthma, rheumatoid arthritis and atopic dermatitis (AD).
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