Overview

  • Product name
    Anti-KDM1 / LSD1 antibody - ChIP Grade
    See all KDM1 / LSD1 primary antibodies
  • Description
    Rabbit polyclonal to KDM1 / LSD1 - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ChIP, IHC-P, IP, ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Human, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 800 to the C-terminus of Human LSD1.

    Read Abcam's proprietary immunogen policy (Peptide available as ab17763.)

  • Positive control
    • Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab17721 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ChIP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 105 kDa (predicted molecular weight: 93 kDa).Can be blocked with Human KDM1 / LSD1 peptide (ab17763).
IHC-Fr Use at an assay dependent concentration. PubMed: 23911933

Target

  • Function
    Histone demethylase that demethylates both 'Lys-4' (H3K4me) and 'Lys-9' (H3K9me) of histone H3, thereby acting as a coactivator or a corepressor, depending on the context. Acts by oxidizing the substrate by FAD to generate the corresponding imine that is subsequently hydrolyzed. Acts as a corepressor by mediating demethylation of H3K4me, a specific tag for epigenetic transcriptional activation. Demethylates both mono- (H3K4me1) and di-methylated (H3K4me2) H3K4me. May play a role in the repression of neuronal genes. Alone, it is unable to demethylate H3K4me on nucleosomes and requires the presence of RCOR1/CoREST to achieve such activity. Also acts as a coactivator of androgen receptor (ANDR)-dependent transcription, by being recruited to ANDR target genes and mediating demethylation of H3K9me, a specific tag for epigenetic transcriptional repression. The presence of PRKCB in ANDR-containing complexes, which mediates phosphorylation of 'Thr-6' of histone H3 (H3T6ph), a specific tag that prevents demethylation H3K4me, prevents H3K4me demethylase activity of KDM1A. Demethylates di-methylated 'Lys-370' of p53/TP53 which prevents interaction of p53/TP53 with TP53BP1 and represses p53/TP53-mediated transcriptional activation. Demethylates and stabilizes the DNA methylase DNMT1. Required for gastrulation during embryogenesis.
  • Tissue specificity
    Ubiquitously expressed.
  • Sequence similarities
    Belongs to the flavin monoamine oxidase family.
    Contains 1 SWIRM domain.
  • Domain
    The SWIRM domain may act as an anchor site for a histone tail.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Amine oxidase (flavin containing) domain 2 antibody
    • Amine oxidase, flavin containing, 2 antibody
    • AOF2 antibody
    • BHC110 antibody
    • BRAF35 HDAC complex protein BHC110 antibody
    • BRAF35-HDAC complex protein BHC110 antibody
    • BRAF35/HDAC complex, 110-kD subunit antibody
    • CPRF antibody
    • EC1 antibody
    • FAD binding protein BRAF35 HDAC complex, 110 kDa subunit antibody
    • Flavin-containing amine oxidase domain-containing protein 2 antibody
    • KDM 1 antibody
    • KDM1 antibody
    • Kdm1a antibody
    • KDM1A_HUMAN antibody
    • KIAA0601 antibody
    • LSD 1 antibody
    • LSD1 antibody
    • Lysine (K) specific demethylase 1 antibody
    • Lysine (K) specific demethylase 1A antibody
    • Lysine demethylase 1A antibody
    • Lysine specific histone demethylase 1 antibody
    • Lysine specific histone demethylase 1A antibody
    • Lysine-specific demethylase 1 antibody
    • Lysine-specific demethylase 1A antibody
    • Lysine-specific histone demethylase 1A antibody
    see all

Images

  • 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 : Anti-KDM1 / LSD1 antibody - ChIP Grade (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

  • 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-iTTM 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).

  • 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.

    See Abreview

  •      ab17721 at a 1/100 dilution in cross-linking ChIP (using formaldehyde) with mouse liver tissue lysate. The protocol from Nguyen, T. T, et al. (2005) Mol. Cell. Biol. 25: 2147-2157 was followed. Semiquantitative PCR was used as the detection step.

    Lane 1: Input

    Lane 2: ab17721

    Lane 3: IgG

    Lane 4: No ab

    This image is courtesy of an Abreview by  Michelle Barton.

    See Abreview

  • 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.

References

This product has been referenced in:
  • Boulding T  et al. LSD1 activation promotes inducible EMT programs and modulates the tumour microenvironment in breast cancer. Sci Rep 8:73 (2018). Read more (PubMed: 29311580) »
  • Cao K  et al. An Mll4/COMPASS-Lsd1 epigenetic axis governs enhancer function and pluripotency transition in embryonic stem cells. Sci Adv 4:eaap8747 (2018). Read more (PubMed: 29404406) »
See all 120 Publications for this product

Customer reviews and Q&As

1-10 of 48 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblast)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
20 µg
Specification
Mouse Embryonic Fibroblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 10 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
20 µg
Specification
HEK293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 10 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (293T)
Gel Running Conditions
Reduced Denaturing (3-8)
Loading amount
1 µg
Specification
293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Oct 02 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa cells)
Permeabilization
Yes - 0.1% TX100, 2 mins
Specification
HeLa cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 02 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Tissue lysate - whole (postnatal day 1 mouse brain)
Total protein in input
500 µg
Immuno-precipitation step
Protein G
Specification
postnatal day 1 mouse brain

Abcam user community

Verified customer

Submitted Mar 15 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
15 µg
Specification
mouse embryonic stem cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Jan 25 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Permeabilization
Yes - Brij
Specification
Brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 0.2% · Temperature: 37°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 21 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (lung)
Loading amount
50 µg
Specification
lung
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Ms. Jihun Lee

Verified customer

Submitted Jan 10 2013

Answer

Thank you for your message and for kindly providing these details.

I look forward to hearing from you with the results of the next experiment, I hope this is successful.

I can suggest it would be important to include an endogenous negative control sample as well if possible.

Should the results be unsatisfactory again, please do not hesitate to let me know and I will be pleased to arrange a free of charge replacement or credit note for you.

Read More

Answer

Thank you for taking the time to contact us and provide this information. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and also gives us vital information for our monitoring of product quality

I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. I would like to reassure you that this antibody is tested and covered by our guarantee for WB and mouse samples.

Before deciding how to proceed with this particular case, I would like to investigate this case in order to understand the problem further. I would also like to offer some suggestions to help optimize the results.

1. I would appreciate if you are able to provide an image which would help me to further understand the problem.

2. Expression of the proteins can change with developmental stage. Could you confirm what expression level you are expecting from the mouse mesenchymal stem cells? Has the expression been confirmed in the samples by other methods?

I can recommend it could be beneficial to consider including a positive control such as Jurkat cells.

3. Has a negative control been included?

4. Has the quality of the sample been assessed using a loading control?

5. How were the mouse mesenchymal stem cells prepared,isolated, and cultured?

6. How were the samples lysed ready for WB? I can recommend to lyse in RIPA lysis buffer which should provide a suitable protein preparation, and to include protease inhibitors.

7. I can recommend to try a lower concentration of antibody to try and reduce the background. 0.5 ug/ml or less.

8. I can suggest to try BSA rather than milk to block. Changing blocking agents can sometimes help to improve results.

9. Is the secondary antibody working well with other primary antibodies?
In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details, including results of the positive and negative control if possible.

Read More

1-10 of 48 Abreviews or Q&A

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