Human RP105/CD180 HEK293 Overexpression Lysate: Product Information
This Human RP105/CD180 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of RP105/CD180 protein (Cat: 11370-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human CD180 (NP_005573.2) extracellular domain (Met 1-Gly 626) was fused with a polyhistidine tag at the C-terminus.
The recombinant human CD180 consists of 614 amino acids and predictes a molecular mass of 69 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhCD180 is approximately 90-100 kDa due to glycosylation.
Human RP105/CD180 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 RP105/CD180 HEK293 Overexpression Lysate: Alternative Names
Human LY64 Overexpression Lysate; Human Ly78 Overexpression Lysate; Human RP105 Overexpression Lysate
RP105/CD180 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. CD18, also known as RP15, is a B-cell surface molecule belonging to the family of pathogen receptors, Toll-like receptors (TLR). CD18 has an extracellular leucine-rich repeats and a short cytoplasmic tail. CD18 / RP15 interact with an extracellular molecule named MD1 and then together form the cell surface receptor complex RP15 / MD1 which induces B-cell activation in humans and mice, leading to proliferation and up-regulation of a costimulatory molecule, B7.2 / CD86. CD18 / RP15 also has a role in LPS response because B cells lacking RP15 show hyporesponsiveness to LPS.
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.
Nagai Y, et al. (2002) Requirement for MD-1 in cell surface expression of RP105/CD180 and B-cell responsiveness to lipopolysaccharide. Journal of the American society of hematology. 99(5): 1699-705.
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