Human TNFRSF1A HEK293 Overexpression Lysate

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Human TNFRSF1A HEK293 Overexpression Lysate: Product Information

Product Description
This Human TNFRSF1A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of TNFRSF1A protein (Cat: 10872-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human TNFRI (NP_001056.1) extracellular domain (Met 1-Thr 211) was fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human TNFRI consists of 201 amino acids after removal of the signal peptide and has a predicted molecular mass of 22.7 kDa. As a result of glycosylation, the apparent molecular mass of rh TNFRI is 30-35 kDa in SDS-PAGE under reducing conditions.

Human TNFRSF1A HEK293 Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
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.
Sample Buffer
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human TNFRSF1A HEK293 Overexpression Lysate: Alternative Names

Human CD120a Overexpression Lysate; Human FPF Overexpression Lysate; Human MS5 Overexpression Lysate; Human p55 Overexpression Lysate; Human p55-R Overexpression Lysate; Human p60 Overexpression Lysate; Human TBP1 Overexpression Lysate; Human TNF-R Overexpression Lysate; Human TNF-R-I Overexpression Lysate; Human TNF-R55 Overexpression Lysate; Human TNFAR Overexpression Lysate; Human TNFR1 Overexpression Lysate; Human TNFR1-d2 Overexpression Lysate; Human TNFR55 Overexpression Lysate; Human TNFR60 Overexpression Lysate

TNFRSF1A 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. CD120a (cluste of differentiation 120a), also known as TNFR1 / TNFRSF1A, is a member of CD family, tumor necrosis factor receptor superfamily. CD120a is one of the most primary receptors for the tumor necrosis factor-alpha. It has been shown to be localized to both plasma membrane lipid rafts and the trans golgi complex with the help of the death domain (DD). CD120a can activate the transcription factor NF-κB, mediate apoptosis, and regulate inflammation processes.
Full Name
tumor necrosis factor receptor superfamily member 1A
References
  • Zola H, et al. (2007) CD molecules 2006-human cell differentiation molecules. J Immunol Methods. 318 (1-2): 1-5.
  • Ho IC, et al. (2009) GATA3 and the T-cell lineage: essential functions before and after T-helper-2-cell differentiation. Nat Rev Immunol. 9 (2): 125-35.
  • Matesanz-Isabel J, et al. (2011) New B-cell CD molecules. Immunology Letters.134 (2): 104-12.
  • Cottin V, et al. (2002) Restricted localization of the TNF receptor CD120a to lipid rafts: a novel role for the death domain. The journal of immunology. 168: 4095-102.
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