Human MD1 HEK293 Overexpression Lysate: Product Information
This Human MD1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of MD1 protein (Cat: 10242-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human LY86 (O95711) (Met1-Ser162) was expressed, fused with the Fc region of human IgG1 at the C-terminus.
The recombinant human LY86/Fc is a disulfide-linked homodimer. The reduced monomer comprises 383 amino acids and has a predicted molecular mass of 42.7 kDa. The apparent molecular mass of the protein is approximately 48 kDa in SDS-PAGE under reducing conditions
Human MD1 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 MD1 HEK293 Overexpression Lysate: Alternative Names
Human dJ80N2.1 Overexpression Lysate; Human MD-1 Overexpression Lysate; Human MD1 Overexpression Lysate; Human MMD-1 Overexpression Lysate
MD1 Background Information
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, dentritic 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 spleen, and also detectable in 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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