Mouse MD1 HEK293 Overexpression Lysate


Mouse MD1 HEK293 Overexpression Lysate: 产品信息

This Mouse MD1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of MD1 protein (Cat: 50062-M08H) from the overexpression lysate was verified.
HEK293 Cells
A DNA sequence encoding the extracellular domain (Met 1-Ser 162) of mouse LY86 (NP_034875.1) precursor was expressed with a C-terminal polyhistidine tag.
The secreted recombinant mouse LY86 consists of 166 amino acids and has a calculated molecular mass of 18 kDa. As a result of glycosylation, the apparent molecualr mass of rmLY86 is approximately 24-30 kDa protein in SDS-PAGE under reducing conditions.

Mouse MD1 HEK293 Overexpression Lysate: 使用指南

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.
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.
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).
稳定性 & 储存条件
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.

Mouse MD1 HEK293 Overexpression Lysate: 别称

Mouse MD-1 Overexpression Lysate; Mouse MD1 Overexpression Lysate

MD1 背景信息

MD-1 and MD-2 are secretory glycoproteins that exist on the cell surface in complexes with transmembrane proteins. MD-1 is anchored by radioprotective 15 (RP15) which is a molecule containing leucine-rich repeats and is expressed on B cells, dendritic cells, and macrophages, while MD-2 is associated with TLR4. MD-1 is required for efficient RP15 cell surface expression and function. It is indicated that the RP15/MD1 complex, in conjunction with TLR4, mediates the innate immune response to LPS in B cells, and also plays a role in protecting against apoptosis, B-cell proliferation, etc. Mouse MD-1 cDNA encodes a 162 amino acid precursor protein with a putative 19 aa signal peptide and two potential N-linked glycosylation sites. It shares 4% and 66% amino acid sequence identity with chicken and human MD-1 respectively. MD-1 is mainly expressed in the spleen, and also detectable in the liver, brain, thymus, and kidney.
lymphocyte antigen 86
  • Miura Y., et al.,(1998), RP105 is associated with MD-1 and transmits an activation signal in human B cells. Blood 92:2815-2822.
  • Begum N.A., et al., (1999), Human MD-1 homologue is a BCG-regulated gene product in monocytes: Its identification by differential display.Biochem. Biophys. Res. Commun. 256:325-329.
  • Mungall A.J., et al.,(2003), The DNA sequence and analysis of human chromosome 6.Nature 425:805-811.
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