Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸ 10 µl 120 614 ₸

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Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18508] to LSD2 / AOF1
Suitable for: Flow Cyt, ICC/IF, IP, WB
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-LSD2 / AOF1 antibody [EPR18508]
See all LSD2 / AOF1 primary antibodies
Description

Rabbit monoclonal [EPR18508] to LSD2 / AOF1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa, HAP1, HCT116, NIH/3T3, C6, RAW 264.7 and PC-12 cell lysates; Human fetal brain and fetal heart lysates; Mouse thymus lysate. ICC/IF: HeLa and A431 cells. IP: K562 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18508
Isotype

IgG
Research areas

Western blot - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

All lanes : Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : KDM1B knockout HeLa cell lysate
Lane 3 : HAP1 cell lysate
Lane 4 : HCT116 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 92 kDa
Observed band size: 95 kDa
why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab193080 observed at 95 kDa. Red - loading control ab8245 observed at 36 kDa.

ab193080 Anti-LSD2 / AOF1 antibody [EPR18508] was shown to specifically react with LSD2 / AOF1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265969 (knockout cell lysate ab258016) was used. Wild-type and LSD2 / AOF1 knockout samples were subjected to SDS-PAGE. ab193080 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling LSD2 / AOF1 antibody (red) with purified ab193080 at a dilution of 1/70. Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.
Western blot - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: LSD2 / AOF1 knockout HAP1 cell lysate (20 µg)
Lane 3: PC12 cell lysate (20 µg)
Lane 4: Raw264.7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab193080 observed at 95 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab193080 was shown to specifically react with LSD2 / AOF1 when LSD2 / AOF1 knockout samples were used. Wild-type and LSD2 / AOF1 knockout samples were subjected to SDS-PAGE. ab193080 and ab8245 (loading control to GAPDH) were diluted at 1/2000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Western blot - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

All lanes : Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080) at 1/2000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 92 kDa
Observed band size: 92 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

All lanes : Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080) at 1/2000 dilution

Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) cell lysate
Lane 2 : Mouse thymus lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 92 kDa
Observed band size: 92 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

All lanes : Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080) at 1/2000 dilution

Lane 1 : C6 (Rat glial tumor cells) cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 92 kDa
Observed band size: 92 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling LSD2 / AOF1 with ab193080 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on HeLa cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab193080 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma) cells labeling LSD2 / AOF1 with ab193080 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on A431 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab193080 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunoprecipitation - Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

LSD2 / AOF1 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab193080 at 1/80 dilution.
Lane 1: K562 whole cell lysate 10ug (Input).
Lane 2: ab193080 IP in K562 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab193080 in K562 whole cell lysate.
Western blot was performed from the immunoprecipitate using ab193080 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)

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