Human GM-CSF Receptor alpha HEK293 Overexpression Lysate

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Human GM-CSF Receptor alpha HEK293 Overexpression Lysate: Product Information

Product Description
This Human GM-CSF Receptor alpha overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of GM-CSF Receptor alpha protein (Cat: 10701-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the extracellular domain (Met 1-Gly 320) of human GM-CSFRα (P15509-1) was expressed with the C-terminal fused polyhistidine tag.
Molecule Mass
The recombinant human GM-CSFRα consists of 309 amino acids and has a predicted molecular mass of 36 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh GM-CSFRα is approximately 60-65 kDa due to glycosylation.

Human GM-CSF Receptor alpha 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 GM-CSF Receptor alpha HEK293 Overexpression Lysate: Alternative Names

Human CD116 Overexpression Lysate; Human CDw116 Overexpression Lysate; Human CSF2R Overexpression Lysate; Human CSF2RAX Overexpression Lysate; Human CSF2RAY Overexpression Lysate; Human CSF2RX Overexpression Lysate; Human CSF2RY Overexpression Lysate; Human GM-CSF-R-alpha Overexpression Lysate; Human GMCSFR Overexpression Lysate; Human GMR Overexpression Lysate; Human SMDP4 Overexpression Lysate

GM-CSF Receptor alpha Background Information

CD116/GM-CSFR has been preferentially associated with M4, M5 subtype of AML but is not specific. The cluster of differentiation (cluster of designation) (often abbreviated as CD) is a protocol used for the identification and investigation of cell surface molecules present on white blood cells initially but found in almost any kind of cell of the body, providing targets for immunophenotyping of cells. Physiologically, CD molecules can act in numerous ways, often acting as receptors or ligands (the molecule that activates a receptor) important to the cell. A signal cascade is usually initiated, altering the behavior of the cell (see cell signaling). Some CD proteins do not play a role in cell signaling, but have other functions, such as cell adhesion. CD116/GM-CSFR is the alpha subunit of the heterodimeric receptor for colony stimulating factor 2, a cytokine which controls the production, differentiation, and function of granulocytes and macrophages. The encoded protein is a member of the cytokine family of receptors. CD116/GM-CSFR is found in the pseudoautosomal region (PAR) of the X and Y chromosomes.
Full Name
colony stimulating factor 2 receptor, alpha, low-affinity (granulocyte-macrophage)
References
  • Sjöblom C, et al. (2002) Granulocyte-macrophage colony-stimulating factor (GM-CSF) acts independently of the beta common subunit of the GM-CSF receptor to prevent inner cell mass apoptosis in human embryos. Biol Reprod. 67(6): 1817-23.
  • Goldstein JI, et al. (2011) Defective leukocyte GM-CSF receptor (CD116) expression and function in inflammatory bowel disease. Gastroenterology. 141(1): 208-16.
  • Saulle E, et al. (2009) Colocalization of the VEGF-R2 and the common IL-3/GM-CSF receptor beta chain to lipid rafts leads to enhanced p38 activation. Br J Haematol. 145(3): 399-411.
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