Anti-HNF1 alpha antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF1 alpha antibody (AB96777)

ab96777, at a 1/100 dilution, staining HNF1 alpha in paraffin embedded A549 Xenograft by Immunohistochemistry.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HNF1 alpha antibody (AB96777)

Immunofluorescence analysis of HepG2 cells fixed in 4% paraformaldehyde at RT for 15 min labelling HNF1 alpha protein at nucleus using ab96777 at a 1/500 dilution (Green). alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody at a 1/1000 dilution (Red).
Scale bar = 10 µM

• IP

Supplier Data

Immunoprecipitation - Anti-HNF1 alpha antibody (AB96777)

Immunoprecipitation of HNF1 alpha protein from HepG2 whole cell extracts using 5 µg of HNF1 alpha antibody [N1N3]. Western blot analysis was performed using HNF1 alpha antibody [N1N3].

All lanes:

Immunoprecipitation - Anti-HNF1 alpha antibody (ab96777) at 5 µg

Lane 1:

HepG2 whole cell extracts

Lane 2:

HepG2 whole cell extracts with control IgG

Lane 3:

HepG2 whole cell extracts (+)

Secondary

All lanes:

EasyBlot HRP-conjugated anti rabbit IgG antibody

Predicted band size: 81 kDa

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• WB

Supplier Data

Western blot - Anti-HNF1 alpha antibody (AB96777)

Samples were separated by 7.5% SDS-PAGE. Corresponding RNA expression data for the same cell lines are based on Human Protein Atlas program.

All lanes:

Western blot - Anti-HNF1 alpha antibody (ab96777) at 1/2000 dilution

Lane 1:

293T whole cell extracts at 30 µg

Lane 2:

A431 whole cell extracts at 30 µg

Lane 3:

HeLa whole cell extracts at 30 µg

Lane 4:

HepG2 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG

Predicted band size: 81 kDa

false

• WB

Lab

Western blot - Anti-HNF1 alpha antibody (AB96777)

Western blot : Anti-HNF1A antibody (ab96777) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab96777 was shown to bind specifically to HNF1A. A band was observed at 75-80 kDa in wild-type A549 cell lysates with no signal observed at this size in HNF1A knockout cell line. To generate this image, wild-type and HNF1A knockout A549 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 then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-HNF1 alpha antibody (ab96777) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

HNF1A knockout A549 cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

U-2 OS cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 75 kDa,80 kDa

false

• WB

Supplier Data

Western blot - Anti-HNF1 alpha antibody (AB96777)

Samples were separated by 7.5% SDS-PAGE.

All lanes:

Western blot - Anti-HNF1 alpha antibody (ab96777) at 1/2000 dilution

All lanes:

Mouse liver tissue extract at 50 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG

Predicted band size: 81 kDa

false