Western blot analysis of extracts from various cell lines using FcRn Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human FcRn protein (hFcRn-Myc/DDK; +), using FcRn Antibody (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
FcRn Antibody recognizes endogenous levels of total FcRn protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly250 of human FcRn protein. Antibodies are purified by peptide affinity chromatography.
The neonatal Fc receptor, FcRn, is an atypical Fc gamma receptor that bears homology to class I MHC (major histocompatibility complex) and associates with β2-microglobulin (1). FcRn is expressed by the vascular endothelium, several epithelial cell types, and hematopoietic cells including several innate immune cell types and B cells (2). FcRn mediates recycling and transcytosis of IgG, resulting in protection of IgG from catabolism and an extended serum half-life of IgG relative to other antibody isotypes (3). In addition, FcRn transports IgG across the placenta and also mediates uptake of maternal IgG from milk, resulting in enhanced humoral immunity in the fetus and neonates (1,4,5). FcRn also plays an important role in initiation of adaptive immunity by transporting immune complexes to intracellular dendritic cell compartments for antigen processing and presentation to T cells (6-8). In addition to IgG, FcRn also binds albumin and similarly protects it from catabolism, leading to extended half-life and high plasma concentrations (9).
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