This Human CD50 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD50 protein (Cat: 10333-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of human ICAM3 (CAA49369.1) (Met 1-His 485) was expressed, with the C-terminal fused polyhistidine tag.
The recombinant human ICAM3 consists of 467 amino acids after removal of the signal peptide and predicts a molecular mass of 50.8 kDa. By SDS-PAGE, the apparent molecular mass of rh ICAM3 is approximately 100-120 kDa due to the glycosylation.
Human CD50 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 CD50 HEK293 Overexpression Lysate: 别称
Human CD50 Overexpression Lysate; Human CDW50 Overexpression Lysate; Human ICAM-3 Overexpression Lysate; Human ICAM-R Overexpression Lysate
The protein ICAM-3, also known as CD50, is a member of the intercellular adhesion molecule (ICAM) family consisting of three members. It is a DC-SIGN ligand that is constitutively expressed on resting leukocytes and is thus an important molecule for the first immune response. ICAM-3 comprises five immunoglobulin-like domains and binds LFA-1 through its two N-terminal domains. It functions not only as an adhesion molecule but also as a potent signaling molecule. ICAM-3 binds to LFA-1 on antigen-presenting cells (APC) stabilizing the T cell-APC interaction, facilitating signaling through the CD3/TCR complex. However, recent evidence using cultured and transformed T cells suggests ICAM-3 may also function in signaling. It has been reported that the CD50 molecule can play a role in developing functionally mature T lymphocytes and its expression increases during the maturation process of T lymphocytes. Also, the interactions of ICAM-3 and LFA-1 facilitate HIV-1- induced virological synapse formation between T cells. ICAM-3 is associated with an increase in cellular radio-resistance and cancer cell proliferation. It could be considered as a candidate for anti-cancer drug development and as a cancer diagnostic marker.
intercellular adhesion molecule 3
Berney SM, et al. (1999) ICAM-3 (CD50) cross-linking augments signaling in CD3-activated peripheral human T lymphocytes. J Leukoc Biol. 65(6): 867-74.
van Buul JD, et al. (2004) ICAM-3 activation modulates cell-cell contacts of human bone marrow endothelial cells. J Vasc Res. 41(1): 28-37.
Sugino H. (2005) ICAM-3, a ligand for DC-SIGN, was duplicated from ICAM-1 in mammalian evolution, but was lost in the rodent genome. FEBS Lett. 579(13): 2901-6.
Park JK, et al. (2010) ICAM-3 enhances the migratory and invasive potential of human non-small cell lung cancer cells by inducing MMP-2 and MMP-9 via Akt and CREB. Int J Oncol. 36(1): 181-92.