Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: KDM1 / LSD1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Jurkat whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab17721 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab17721 was shown to specifically react with KDMI/LSD1 in wild-type HAP1 cells. No band was observed when KDMI/LSD1 knockout samples were examined.

ICC/IF image of ab17721 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab17721, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

All lanes :

Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates at 20 µg
Lane 2 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 µg
Lane 4 : C2C12 (Mouse myoblasts myoblast) whole cell lysates
Lane 5 : Mouse skeletal muscle lysates at 20 µg
Lane 6 : C6 (Rat glial tumor glial cell) whole cell lysates at 20 µg

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 93 kDa


Exposure time: 50 seconds

Observed MW: 110kd

Blocking Buffer & Diluting buffer concentration: 5% NFDM/TBST

All lanes : Anti-KDM1/LSD1 antibody - Nuclear Marker (ab17721) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat whole cell lysate (ab7899)
Lane 3 : NIH/3T3 whole cell lysate (ab7179)

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution

Performed under reducing conditions.

Predicted band size: 93 kDa

ab17721 staining LSD1 in the mouse c2c12 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 2% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/200) for 1 hour at 25°C. A diluted (1/2000) Alexa Fluor® 568-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

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Human 293T cells fixed in 4% paraformaldehyde were immunostained with ab17721 (1/200) for LSD1 and detected using FITC labelled anti-rabbit (Green). Nuclear DNA is stained blue with DAPI.