Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human AP4B1 protein (hAP4B1-Myc/DDK; +), using AP4B1 Antibody (upper) and Myc-Tag (71D10) Rabbit mAb #2278 (lower).
Western blot analysis of extracts from various cell lines using AP4B1 Antibody (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. 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
AP4B1 Antibody recognizes endogenous levels of total human AP4B1 protein. Weak reactivity has been observed in rodent.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ile708 of human AP4B1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
AP (adaptor protein) complexes regulate intracellular membrane trafficking along endocytic and secretory pathways (1). There are five AP complexes (AP1-5) that exist as heterotetrameric complexes functioning at distinct intracellular compartments. The AP4 complex, which includes AP4E1, AP4B1, AP4M1, and AP4S1, functions at the trans-Golgi network (TGN) (2-4). AP-4 associates with Atg9A which is critical in autophagosome biogenesis (5). Deficiency in AP-4 led to Atg9A retention within the trans-Golgi network with defective axonal autophagosome generation (5). AP-4 deficiency has been linked to a group of neurodegenerative diseases known as hereditary spastic paraplegia. Mutation in the AP4B1 gene was identified as a cause for the neurological disorder hereditary spastic paraplegia type 47 (SPG47) (6,7).
Explore pathways + proteins related to this product.
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