Western blot analysis of extracts from various cell lines using ARL8B Antibody (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Decreased signal of ARL8B in OVCAR-3 cells is predicted from RNAseq data and confirms the specificity of the antibody for ARL8B.
Western blot analysis of extracts from 293T cells, untransfected (-) or transfected (+) with constructs expressing Myc/DDK-tagged full-length human ARL8B protein (hARL8B-Myc/DDK) or untagged human ARL8A protein (hARL8A), using ARL8B Antibody (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
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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
ARL8B Antibody recognizes endogenous levels of total ARL8B protein. This antibody does not cross-react with ARL8A.
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala137 of human ARL8B protein. Antibodies are purified by protein A and peptide affinity chromatography.
The small GTPases of the Arf and Rab families have critical roles in cytoskeletal organization and membrane/vesicular transport (1,2). The related ADP-ribosylation factor-like (ARL) family member ARL8, including two paralogs ARL8A and ARL8B, has emerged as an important regulator of lysosomal transport to the cell periphery (3,4). Lysosome positioning is a critical determinant of a number of physiological processes including plasma membrane repair, cell migration, antigen presentation, and response to nutrient availability. Membrane association of ARL8B is regulated by the BLOC-one related complex (BORC), a multi-subunit complex under the control of the mTOR pathway (5,6). Effectors of GTP-bound ARL8B include PLEKHM2/SKIP and the Vps41 subunit of the HOPS complex (7). Studies have found that ARL8B may be a critical factor for NK-mediated cytotoxicity by driving polarization of lytic granules toward the immune synapse (8). In addition, ARL8B has been shown to act as a negative regulator of autophagy (9). Knockdown of ARL8B resulted in increased fusion of autophagosomes with lysosomes. Furthermore, ARL8B depleted cells had reduced levels of pathogenic substrates associated with neurodegenerative diseases.
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