Anti-PTP1B antibody [EP1841Y]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PTP1B antibody [EP1841Y] (AB75856)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling PTP1B with purified ab75856 at 1/500 dilution (5.3 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL) was used as the secondary antibody only control.

• WB

Lab

Western blot - Anti-PTP1B antibody [EP1841Y] (AB75856)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PTP1B (phospho S378) antibody [EP1840Y] (<a href='/en-us/products/primary-antibodies/ptp1b-phospho-s378-antibody-ep1840y-ab76239'>ab76239</a>) at 1/10000 dilution

Lane 1:

Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

HeLa treated with 100ng/ml Calyculin A for 10mins whole cell lysate at 15 µg

Lane 3:

HeLa treated with 100ng/ml Calyculin A for 10mins whole cell lysate, then the membrane treated with phosphatase at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 50 kDa

false

Exposure time: 20s

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PTP1B antibody [EP1841Y] (AB75856)

ab75856, at 1/50 dilution, staining PTP1B in human tonsil, by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PTP1B antibody [EP1841Y] (AB75856)

Intracellular Flow Cytometry analysis of Jurkat cells labelling PTP1B with ab75856 at 1/2000 (red). Cells were fixed with 90% methanol. ab150077, Alexa Fluorr^® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG (ab172730). Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

• WB

Unknown

Western blot - Anti-PTP1B antibody [EP1841Y] (AB75856)

All lanes:

Western blot - Anti-PTP1B antibody [EP1841Y] (ab75856) at 1/2000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/1000 dilution

Predicted band size: 50 kDa

Observed band size: 50 kDa

false

• WB

Supplier Data

Western blot - Anti-PTP1B antibody [EP1841Y] (AB75856)

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 (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-PTP1B antibody [EP1841Y] (ab75856)

Predicted band size: 50 kDa

false