Human CD89 HEK293 Overexpression Lysate: Product Information
This Human CD89 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD89 protein (Cat: 10414-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the pro form of human FCAR extracellular domain (NP_001991.1) (Met 1-Asn 227) was expressed with a C-terminal polyhistidine tag.
The secreted recombinant human CD89 is a monomeric protein consisting of 217 amino acids and predicts a molecular mass of 25 kDa. The apparent molecular mass of rhCD89 is approximately 45-50 kDa in SDS-PAGE under reducing conditions.
Human CD89 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 CD89 HEK293 Overexpression Lysate: Alternative Names
Human CD89 Overexpression Lysate; Human CTB-61M7.2 Overexpression Lysate; Human FcalphaRI Overexpression Lysate; Human FCAR Overexpression Lysate; Human XXbac-BPG230H20.5 Overexpression Lysate
CD89 Background Information
FCAR, also called FcαRI or CD89, is a type I transmembrane receptor for Fc region of IgA which is the most abundant immunoglobulin in mucosal areas but is only the second most common antibody isotype in serum. This receptor is present on the surface of myeloid lineage cells such as neutrophils, monocytes, macrophages, and eosinophils, especially phagocytes located in mucosal areas. Upon ligand IgA binding, FcαRI associates with the FcR γ signaling molecule bearing the immunoreceptor tyrosine-based activation motif (ITAM) through a unique charge-based mechanism and triggers multiple cell-mediated immune responses. It has been reported that Fc RI is a dual-function receptor that can mediate both inflammatory and anti-inflammatory responses depending on the type of interaction with its ligand. Sustained aggregation of FCAR results in activation of target-cell functions such as antigen presentation and cytokine release. In contrast, Monomeric targeting with serum IgA or with a variety of anti-FcαRI Fab fragments triggers an inhibitory response and additionally induces apoptosis. FcαRI thus play an fundamental role in preventing tumor development and growth, as well as in controlling inflammation.
Fc fragment of IgA receptor
Maliszewski, C.R. et al., 1990, J. Exp. Med. 172: 1665-1672.
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