|H M R||Endogenous||160||Rabbit IgG|
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
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 November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised June 2016
Protocol Id: 263
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.
PTPN14 (D5T6Y) Rabbit mAb recognizes endogenous levels of total PTPN14 protein.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu737 of human PTPN14 protein.
Tyrosine-protein phosphatase non-receptor type-14 (PTPN14, Pez, PTPD2 and PTP36) is an evolutionarily conserved non-membrane tyrosine phosphatase with homology to the band 4.1 family of proteins (1-3). The PTPN14 protein contains an amino-terminal FERM (4.1-ezrin-radixin-moesin) domain, which suggests plasma membrane localization of the protein, and a carboxy-terminal protein tyrosine phosphatase (PTP) domain (4). Research studies have identified possible roles for PTPN14 in multiple, diverse signaling pathways, including cell growth and proliferation, cell migration and adhesion, and development. The PTPN14 phosphatase regulates the subcellular localization of YAP in a cell density-dependent manner, indicating a role for PTPN14 in the Hippo signaling pathway (5). The Drosophila PTPN14 homolog Pez localizes to adherens junctions, where it may regulate cell motility through dephosphorylation of β-catenin (3). PTPN14 may play a role in epithelial-mesenchymal transition through effects on the TGF-β signaling pathway (6), and interacts with VEGFR3, a receptor tyrosine kinase involved in lymphangiogenesis (7). Loss-of-function mutations in the PTPN14 gene are associated with colorectal cancer (8), and choanal atresia and lymphedema, an autosomal recessive disorder characterized by defects in both nasal passage development and lymphangiogenesis (7).
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|13808S||100 µl||$ 260|