Anti-KDM1/LSD1 抗体 - Nuclear Marker

• WB

Lab

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

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.

All lanes:

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (ab17721)

Predicted band size: 92 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

ab17721 staining KDM1 / LSD1 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 overnight at 4°C with ab17721 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

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.

This image is courtesy of Katia Ancelin

• ICC/IF

AbReview22170****

Immunocytochemistry/ Immunofluorescence - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

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.

Image courtesy of anonymous abreviewer

• WB

Lab

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

Observed MW : 110kd

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

All lanes:

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (ab17721) at 1/1000 dilution

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

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 (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 92 kDa

Observed band size: 110 kDa

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Exposure time: 50s

• WB

Project

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

All lanes:

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (ab17721) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

Western blot - Jurkat whole cell lysate (<a href='/products/cell-lysates/jurkat-whole-cell-lysate-ab7899'>ab7899</a>) at 20 µg

Lane 3:

Western blot - NIH/3T3 whole cell lysate (<a href='/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 20 µg

Secondary

All lanes:

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

Predicted band size: 92 kDa

false

• WB

CiteAb

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

Western Blotting using Anti-KDM1/LSD1 antibody - Nuclear Marker, ab17721. Publication image from Chen, R. et al., 2019, Nat Commun, 30894540. Legend direct from paper.

GFI1B recruits LSD1 to β-Catenin containing complexes. a, b Western blot analysis of indicated proteins after immune precipitation of endogenous β-catenin from 293T cells transfected with the indicated GFI1B constructs. c GST-E-Cadherin pull down of endogenous β-catenin in U2OS cells expressing inducible GFI1B (Tet-ON system) followed by Western blot analysis. d Dot Plot showing BioID interactions of β-catenin-BirA*-Flag coexpressing an Empty Vector (1), the wild-type form of GFI1B (2), GFI1B lacking the SNAG domain (3) or GFI1B lacking the WRD domain (4) with the indicated Wnt pathway regulators in Flp-In T-REx HEK293 cells. e Heat map illustrating biological processes and molecular functions of associated GO terms. The enrichment score of each GO term is shown as the −log10 of corrected P values, indicated by different color intensities

false

• WB

CiteAb

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

Western Blotting using Anti-KDM1/LSD1 antibody - Nuclear Marker, ab17721. Publication image from Chen, R. et al., 2019, Nat Commun, 30894540. Legend direct from paper.

GFI1B recruits LSD1 to β-Catenin containing complexes. a, b Western blot analysis of indicated proteins after immune precipitation of endogenous β-catenin from 293T cells transfected with the indicated GFI1B constructs. c GST-E-Cadherin pull down of endogenous β-catenin in U2OS cells expressing inducible GFI1B (Tet-ON system) followed by Western blot analysis. d Dot Plot showing BioID interactions of β-catenin-BirA*-Flag coexpressing an Empty Vector (1), the wild-type form of GFI1B (2), GFI1B lacking the SNAG domain (3) or GFI1B lacking the WRD domain (4) with the indicated Wnt pathway regulators in Flp-In T-REx HEK293 cells. e Heat map illustrating biological processes and molecular functions of associated GO terms. The enrichment score of each GO term is shown as the −log10 of corrected P values, indicated by different color intensities

false

• WB

CiteAb

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

Western Blotting using Anti-KDM1/LSD1 antibody - Nuclear Marker, ab17721. Publication image from Chen, R. et al., 2019, Nat Commun, 30894540. Legend direct from paper.

GFI1B recruits LSD1 to β-Catenin containing complexes. a, b Western blot analysis of indicated proteins after immune precipitation of endogenous β-catenin from 293T cells transfected with the indicated GFI1B constructs. c GST-E-Cadherin pull down of endogenous β-catenin in U2OS cells expressing inducible GFI1B (Tet-ON system) followed by Western blot analysis. d Dot Plot showing BioID interactions of β-catenin-BirA*-Flag coexpressing an Empty Vector (1), the wild-type form of GFI1B (2), GFI1B lacking the SNAG domain (3) or GFI1B lacking the WRD domain (4) with the indicated Wnt pathway regulators in Flp-In T-REx HEK293 cells. e Heat map illustrating biological processes and molecular functions of associated GO terms. The enrichment score of each GO term is shown as the −log10 of corrected P values, indicated by different color intensities

false

• WB

CiteAb

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

Western Blotting using Anti-KDM1/LSD1 antibody - Nuclear Marker, ab17721. Publication image from Chen, R. et al., 2019, Nat Commun, 30894540. Legend direct from paper.

GFI1B interacts with β-catenin and several regulators of the Wnt/β-catenin signaling pathway. a Immune precipitation (IP) with anti Flag antibody from 293T cells overexpressing GFI1B-Flag followed by mass spectrometric analysis. b Dot Plot showing BioID interactions of GFI1B-BirA*-Flag with the indicated Histone modifying enzymes and Wnt pathway regulators in Flp-In T-REx HEK293 cells. Node color represents the average spectral counts. The edge color depicts the confidence score of the interaction (BFDR ≤ 1% as high confidence, 1% < BFDR ≤ 5% as medium confidence or 5% < BFDR as low confidence score). The relative abundance of prey across the bait is represented by the size of the circle. c A previously described Wnt regulatory domain (WRD) is present in GFI1B28. The indicated Flag-tagged and mutated forms used for IP. d, e Immune precipitation with anti Flag antibodies from 293T cells overexpressing WT or the indicated mutated forms of GFI1B followed by western blot. f–h Immune precipitation with an anti GFI1B antibody from K562 (f), HEL (g), and CHIR99021 treated K562 (h) cells

false

• WB

CiteAb

Western blot - Anti-KDM1/LSD1 antibody - Nuclear Marker (AB17721)

Western Blotting using Anti-KDM1/LSD1 antibody - Nuclear Marker, ab17721. Publication image from Chen, R. et al., 2019, Nat Commun, 30894540. Legend direct from paper.

GFI1B interacts with β-catenin and several regulators of the Wnt/β-catenin signaling pathway. a Immune precipitation (IP) with anti Flag antibody from 293T cells overexpressing GFI1B-Flag followed by mass spectrometric analysis. b Dot Plot showing BioID interactions of GFI1B-BirA*-Flag with the indicated Histone modifying enzymes and Wnt pathway regulators in Flp-In T-REx HEK293 cells. Node color represents the average spectral counts. The edge color depicts the confidence score of the interaction (BFDR ≤ 1% as high confidence, 1% < BFDR ≤ 5% as medium confidence or 5% < BFDR as low confidence score). The relative abundance of prey across the bait is represented by the size of the circle. c A previously described Wnt regulatory domain (WRD) is present in GFI1B28. The indicated Flag-tagged and mutated forms used for IP. d, e Immune precipitation with anti Flag antibodies from 293T cells overexpressing WT or the indicated mutated forms of GFI1B followed by western blot. f–h Immune precipitation with an anti GFI1B antibody from K562 (f), HEL (g), and CHIR99021 treated K562 (h) cells

false