HRP Anti-SHP1 antibody [Y476]

• WB

Lab

Western blot - HRP Anti-SHP1 antibody [Y476] (AB205427)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab205427 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - HRP Anti-SHP1 antibody [Y476] (ab205427) at 1/5000 dilution

All lanes:

Jurkat (Human) Whole Cell Lysate at 10 µg

Predicted band size: 68 kDa

Observed band size: 65 kDa

true

Exposure time: 90s

• WB

Lab

Western blot - HRP Anti-SHP1 antibody [Y476] (AB205427)

False colour image of Western blot : HRP Anti-SHP1 antibody [Y476] staining at 1/5000 dilution, shown in black; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab205427 was shown to bind specifically to SHP1. A band was observed at 70 kDa in wild-type THP-1 cell lysates with no signal observed at this size in PTPN6 knockout cell line ab273717 (knockout cell lysate ab273771). To generate this image, wild-type and PTPN6 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with Optiblot (ECL reagent ab133456) and imaged with 2 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - HRP Anti-SHP1 antibody [Y476] (ab205427) at 1/5000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

PTPN6 knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN6 knockout THP-1 cell line (<a href='/ko/products/cell-lines/human-ptpn6-knockout-thp-1-cell-line-ab273717'>ab273717</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 68 kDa

Observed band size: 70 kDa

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