Anti-Heme Oxygenase 1 antibody [1F12-A6]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [1F12-A6] (AB223349)

100% methanol fixed HaCaT (human keratinocyte cell line) cells stained for Heme Oxygenase 1 (green) using ab223349 at a 1/100 dilution in ICC/IF. Secondary : FITC Goat Anti-Mouse at 1/50 dilution for 1 hour at RT.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [1F12-A6] (AB223349)

2% formaldehyde fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Heme Oxygenase 1 (green) using ab223349 at a 1/100 dilution in ICC/IF. Secondary : FITC Goat Anti-Mouse at 1/200 dilution for 2 hours at RT. Counterstain : DAPI (blue) nuclear stain at 1/40000 dilution for 2 hours at RT.

(A) DAPI nuclear stain (B) ab223349 (C) Merge.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Heme Oxygenase 1 antibody [1F12-A6] (AB223349)

2% formaldehyde fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for Heme Oxygenase 1 (yellow) using ab223349 at a 1/100 dilution in ICC/IF. Secondary : R-PE Goat Anti-Mouse at 1/200 dilution for 2 hours at RT. Counterstain : DAPI (blue) nuclear stain at 1/40000 dilution for 2 hours at RT.

(A) DAPI nuclear stain (B) ab223349 (C) Merge.

• IHC-P

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Immunohistochemistry paraffin embedded sections - Anti-Heme Oxygenase 1 antibody [1F12-A6] (AB223349)

Bouin's fixative, paraffin embedded mouse back skin tissue stained for Heme Oxygenase 1 (green) with ab223349 at a 1/100 dilution in immunohistochemical analysis. Secondary Antibody : FITC Goat Anti-Mouse at 1/50 for 1 hour at RT.

• WB

Supplier Data

Western blot - Anti-Heme Oxygenase 1 antibody [1F12-A6] (AB223349)

Blocking : 1.5% BSA for 30 minutes at RT.

All lanes:

Western blot - Anti-Heme Oxygenase 1 antibody [1F12-A6] (ab223349) at 1/1000 dilution

All lanes:

HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 15 µg

Secondary

All lanes:

Sheep Anti-Mouse IgG: HRP

Predicted band size: 33 kDa

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• WB

Lab

Western blot - Anti-Heme Oxygenase 1 antibody [1F12-A6] (AB223349)

False colour image of Western blot : Anti-Heme Oxygenase 1 antibody [1F12-A6] staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab223349 was shown to bind specifically to Heme Oxygenase 1. A band was observed at 33 kDa in wild-type A549 cell lysates with no signal observed at this size in HMOX1 knockout cell line ab269503 (HMOX knockout A549 lysate ab259782). To generate this image, wild-type and HMOX1 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

All lanes:

Western blot - Anti-Heme Oxygenase 1 antibody [1F12-A6] (ab223349) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

HMOX1 knockout A549 cell lysate at 20 µg

Lane 3:

Mouse Spleen cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Lane 5:

HEK-293 cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 33 kDa

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