Antibodies, Proteins, Kits and Reagents for Life Science

Anti-KDM1/LSD1 antibody [1B2F2] (ab69535)

Key features and details

Mouse monoclonal [1B2F2] to KDM1/LSD1
Suitable for: ICC/IF, WB
Knockout validated
Reacts with: Mouse, Human
Isotype: IgG1

Product name

Anti-KDM1/LSD1 antibody [1B2F2]
See all KDM1/LSD1 primary antibodies
Description

Mouse monoclonal [1B2F2] to KDM1/LSD1
Host species

Mouse
Tested Applications & Species

Application Species
ICC/IF
Human

WB
Mouse

Human

See all applications and species data
Immunogen

Recombinant human LSD1 truncated protein
Positive control
- WB: NIH/3T3, Jurkat, HeLa and HAP1 cell lysates. ICC/IF: HAP1-KDM1A cells.
General notes

Concentration: See product label
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Application	Species
ICC/IF	Human
WB	Mouse Human

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.09% Sodium azide
Constituents: 50% Glycerol, PBS
Concentration information loading...
Purity

Protein G purified
Clonality

Monoclonal
Clone number

1B2F2
Isotype

IgG1
Research areas

Western blot - Anti-KDM1/LSD1 antibody [1B2F2] (ab69535)

All lanes : Anti-KDM1/LSD1 antibody [1B2F2] (ab69535)

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : KDM1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 93 kDa
Observed band size: 110 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab69535 observed at 110 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

ab69535 was shown to react with KDM1/LSD1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265790 (knockout cell lysate ab256965) was used. Wild-type HeLa and KDM1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab69535 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-KDM1 / LSD1 antibody [1B2F2] (ab69535)

ab69535 staining KDM1A in wild-type HAP1 cells (top panel) and KDM1A knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab69535 at 1μg/ml concentration and ab202272 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Western blot - Anti-KDM1 / LSD1 antibody [1B2F2] (ab69535)

All lanes : Anti-KDM1/LSD1 antibody [1B2F2] (ab69535) at 1/1000 dilution

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : KDM1/LSD1 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 93 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab69535 observed at 110 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab69535 was shown to specifically react with KDM1/LSD1 in wild-type HAP1 cells. No band was observed when KDM1/LSD1 knockout samples were used. Wild-type and KDM1/LSD1 knockout samples were subjected to SDS-PAGE. ab69535 and ab181602 (loading control to GAPDH) were diluted to 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.
Western blot - Anti-KDM1 / LSD1 antibody [1B2F2] (ab69535)

All lanes : Anti-KDM1/LSD1 antibody [1B2F2] (ab69535) at 1 µg/ml

Lane 1 : Molecular weight marker
Lane 2 : HeLa cell extract
Lane 3 : NIH/3T3 cell extract

Developed using the ECL technique.

Predicted band size: 93 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
Additional bands at: 112 kDa. We are unsure as to the identity of these extra bands.

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