The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/2000. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
1/50 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Application notesIs unsuitable for IP.
FunctionMay play an important role in CKII- and p60c-src-induced signal transduction cascades.
Sequence similaritiesBelongs to the protein-tyrosine phosphatase family. Non-receptor class 1 subfamily. Contains 1 tyrosine-protein phosphatase domain.
Post-translational modificationsOxidized on Cys-215; the Cys-SOH formed in response to redox signaling reacts with the alpha-amido of the following residue to form a 4-amino-3-isothiazolidinone serine cross-link, triggering a conformational change that inhibits substrate binding and activity. The active site can be restored by reduction.
ab75856, at 1/50 dilution, staining PTP1B in human tonsil, by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue.
Western blot - Anti-PTP1B antibody [EP1841Y] (ab75856)
Predicted band size : 50 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: PTPB1 knockout HAP1 cell lysate (20 µg) Lane 3: Jurkat cell lysate (20 µg) Lane 4: MCF7 cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab75856 observed at 54 kDa. Red - loading control, ab8245, observed at 37 kDa. ab75856 was shown to specifically react with PTP1B when PTP1B knockout samples were used. Wild-type and ProteinX knockout samples were subjected to SDS-PAGE. ab75856 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Western blot - PTP1B antibody [EP1841Y] (ab75856)
All lanes : Anti-PTP1B antibody [EP1841Y] (ab75856) at 1/2000 dilution
Lane 1 : HeLa cell lysate Lane 2 : HepG2 cell lysate Lane 3 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat anti-rabbit HRP at 1/1000 dilution
Predicted band size : 50 kDa Observed band size : 50 kDa
References for Anti-PTP1B antibody [EP1841Y] (ab75856)