This Human CD1D overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD1D protein (Cat: 11009-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of human CD1D (NP_001757.1) (Met 1-Ser 301) was expressed, with a C-terminal polyhistidine tag.
The recombinant human CD1D consists of 293 amino acids and predicts a molecular mass of 33.3 kDa. As a result of glycosylation, rh CD1D migrates as an approximately 45-50 kDa band in SDS-PAGE under reducing conditions.
Human CD1D 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.
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.
Human CD1D HEK293 Overexpression Lysate: 别称
Human CD1A Overexpression Lysate; Human CD1D Overexpression Lysate; Human MGC34622 Overexpression Lysate; Human R3 Overexpression Lysate
The cluster of differentiation (CD) system is commonly used as cell markers in Immunophenotyping. Different kinds of cells in the immune system can be identified through the surface CD molecules associating with the immune function of the cell. There are more than 320 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. Cluster of differentiation 1 (CD1) is a member of the CD system. It's a family of glycoproteins expressed on the surface of various human antigen-presenting cells which are implicated in the presentation of lipid antigens to T-cells. Due to the different lipid anchoring, the CD1 family is classified into two groups: group1 (CD1a-c) and group2 (CD1d). CD1d with lipid antigens activate NK T-cells which rapidly produce Th1 and Th2 cytokines after been activated.
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.
Joyce S. (2001) CD1d and natural T cells: how their properties jump-start the immune system. Cell Mol Life Sci. 58 (3): 442-69.