Western blot analysis of extracts from various cell lines using PTPRF/LAR (E8W3H) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
|MW (kDa)||150, 213|
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
PTPRF/LAR (E8W3H) Rabbit mAb recognizes endogenous levels of total PTPRF/LAR protein. This antibody detects the E-subunit of the processed PTPRF/LAR protein. Weak reactivity is seen in mouse and rat.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala1109 of human PTPRF/LAR protein.
Receptor type protein tyrosine phosphatase F (PTPRF, LAR) is a transmembrane PTP that helps to regulate insulin signaling, cell proliferation and cell migration. The PTPRF protein is composed of an extracellular segment that contains several Ig-like and fibronectin (Fn-III) domains, a transmembrane region and a pair of cytoplasmic phosphatase domains (1,2). Functional studies reveal that the membrane-associated D1 phosphatase domain is responsible for substrate dephosphorylation, while the D2 domain is important for substrate specificity (3). PTPRF negatively regulates insulin signaling through dephosphorylation of insulin receptor and insulin receptor substrate (4). This phosphatase activates the pro-apoptotic DAPK serine/threonine kinase by removing a phosphate at Tyr491/492, while the kinase Src replaces the phosphate to inactivate DAPK at the same time it down regulates PTPRF expression (5). PTPRF is commonly found at focal adhesions where it interacts with liprin, which localizes the phosphatase to the membrane, and the Rac/Rho family GTPase Trio (6). Localization of PTPRF at adherens junctions results in PTPRF modification of β-catenin, which inhibits cell migration by limiting the amount of available cytosolic β-catenin (7).
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