Anti-Ferritin Heavy Chain antibody

• WB

Unknown

Western blot - Anti-Ferritin Heavy Chain antibody (AB65080)

All lanes:

Western blot - Anti-Ferritin Heavy Chain antibody (ab65080) at 1 µg/mL

Lane 1:

Western blot - Recombinant Human Ferritin Heavy Chain protein (Tag Free) (<a href='/en-us/products/proteins-peptides/recombinant-human-ferritin-heavy-chain-protein-ab78877'>ab78877</a>) at 0.1 µg

Lane 2:

Western blot - Recombinant Human Ferritin Heavy Chain protein (Tag Free) (<a href='/en-us/products/proteins-peptides/recombinant-human-ferritin-heavy-chain-protein-ab78877'>ab78877</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 21 kDa

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Exposure time: 30s

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ferritin Heavy Chain antibody (AB65080)

IHC image of ab65080 staining in Human Normal Liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab65080, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Unknown

Western blot - Anti-Ferritin Heavy Chain antibody (AB65080)

Ferritin Heavy Chain contains a number of potential phosphorylation sites (SwissProt data) which may explain it running at a slightly higher than predicted molecular weight (24 kDa).

All lanes:

Western blot - Anti-Ferritin Heavy Chain antibody (ab65080) at 1 µg/mL

All lanes:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa,24 kDa

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Exposure time: 3min

• WB

Lab

Western blot - Anti-Ferritin Heavy Chain antibody (AB65080)

Western blot : Anti-FTH1 antibody (ab65080) staining at 1 ug/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab65080 was shown to bind specifically to FTH1. A band was observed at 21 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in FTH1 knockout cell line. To generate this image, wild-type and FTH1 knockout HEK-293T 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-Ferritin Heavy Chain antibody (ab65080) at 1 µg/mL

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

FTH1 knockout HEK-293T cell lysate at 20 µg

Lane 3:

Wild-type MCF7 ab269599 cell lysate at 20 µg

Lane 4:

FTH1 knockout MCF7 ab269493 cell lysate at 20 µg

Lane 5:

SH-SY5Y 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: 21 kDa

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