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 133 Publications for this product

Customer reviews and Q&As

1-10 of 27 Q&A

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.

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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.

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Answer

Thank you for contacting Abcam regarding ab17721.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement from a new lot.

To check the status of the order please contact our Customer Service team and reference this number.

In the meantime, would it be possible to provide the details of your protocol? I will then forward this on to our laboratory for further investigation. Thank you in advance for your cooperation.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number ***.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with a different lot of ab17721.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

Thanks for your reply.

I have requested the immunogen information used to produce ab90966. I will forward it to you as soon as I receive the details.

We typically only provide the discount testing program codes prior to purchase of the product to be tested. However I can make an exception in this case as you were not aware of this promotion. The discount codeassociated with this promotion will provide you one free primary antibody of your choice after you have submitted your Abreview. Please see below for your discount code information, including expiration date of the code:

=======

DISCOUNT CODE: 100%ABR-T71FL
Expiration date: September 30 2012

I am very pleased to hear you would like to accept our offer and test ab90966 in Toxoplasma. This code will give you: 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for Toxoplasma and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for your enquiry.

We do not have small sample sizes available. In lieu of this, we do have a testing program which you may be intersted in. I have described it below using ab90966 as an example. But the promotion could be applied to any of the antibodies you have mentioned here. Please let me know if you would like to take advantage of the promotion with 1 or more antibodies or if you have any questions.

======

Thank you very much for your interest in ab90966.

To our knowledge, ab90966 has not been tested in Toxoplasma. Therefore, I can offer a discount off a future purchase if you buy ab90966 now, test it in Toxoplasma and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody. If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab90966 in Toxoplasma Iwill then send a discount code. This code must be issued before purchasing ab90966 so please wait for my reply before ordering.

2. Purchase ab90966 either by phone, fax, or online (www.abcam.com).

3. Test it in Toxoplasma.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab90966 turns out to be unsuitable forToxoplasma you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

I am sorry that the protocol suggestions made have not lead to a significant improvement in the non-specificity observed. It does however appear that the non-specificity has been reduced. I would suggest to your customer that it would be worthile trying to optimise the protocol further, for example using a higher dilution of the antibody (1/2000) as well as reducing the amount of protein loaded on the gel (to 10 ug/well). This may lead to further improvements in the results observed.

I can however understand the frustrations of your customer and the time and effort they ahve put into optimising the results with this antibody. I ahve therefore generated a credit note as requested. The credit note ID is xxxxx.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

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Question

Dear technical support team:

This customer has purchased ab17721 (Anti-KDM1 / LSD1 antibody - ChIP Grade) and has conducted the wb several times with mouse sample. The results show no major band and high background, therefore, she wants to ask for your help to modify her experiment step, could you please offer any suggestion to help her?

I also attached her image in this letter and her experiment step as follow:
1. Order details:
Batch number: GR50119-1
Po:
Abcam product code: ab17721
Antibody storage conditions (temperature/reconstitution etc) -20oC
2. Please describe the problem (high background, wrong band size, more bands, no band etc). More bands, no specific band
3. On what material are you testing the antibody in WB?
· Species: Human
· What’s cell line or tissue: K562, HL60, U937, and Jurkat…..
· Cell extract or Nuclear extract: Cell extract
· Purified protein or Recombinant protein: purified
3. The lysate
How much protein was loaded: 20-30 µg
What lysis buffer was used: RIPA
What protease inhibitors were used: inhibitor cocktails
What loading buffer was used: 4X sample buffer (1M Tris-HCl pH 6.8, SDS, Glycerol, BPB)
Phosphatase inhibitors : sodium fluride, sodium orthovanadate
Did you heat the samples: temperature and time: 95oC 3-5 mins
4. Electrophoresis/Gel conditions/ Transfer conditions
Reducing or non reducing gel: Reducing
Reducing agent: BME
Gel percentage : 8% SDS PAGE
Transfer conditions: (Type of membrane, Protein transfer verified): PVDF, pre-stained marker
5. Blocking conditions
Buffer: TBST
Blocking agent: 5% milk
Incubation time: 1-2h
Incubation temperature: RT
6. Primary Antibody
Species:HumanReacts against: KDM1/LSD1
· At what dilution(s) have you tested this antibody:1/1000
· What dilution buffer was used: TBST in 5% Milk
· Incubation time: O/N
· Incubation temperature: 4oC
· What washing steps were done: more than three times using TBST buffer
7. Secondary Antibody
Species:Reacts against: Rabbit IgGAt what dilution(s) have you tested this antibody: 1/5000-1/10000Incubation time: 45-60 minutes
Wash steps: 3 times, 10 minutes each time, TBST buffer
Fluorochrome or enzyme conjugate: enzyme conjugate
Do you know whether the problems you are experiencing come from the secondary? No
8. Detection method
ECl, ECl+, other detection method: ECl
9. Did you apply positive and negative controls along with the samples? Please specify. No
10. Optimization attempts
· How many times have you tried the Western? 3 times
· Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): Yes
· Do you obtain the same results every time e.g. are background bands always in the same place? Yes
· What steps have you altered? Higher dilution of 2Ab
Could you please help this customer to solve the problem?
Thanks for your kindly help
Best regards

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Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab17721. This antibody has been used in house with 5% BSA used as the blocking agent. I would therefore suggest that you try thins blocking agent, in TBST to block the membrane, as well as in the diluent of the primary and secondary antibody. This change from milk blocking to BSA blocking can sometimes have a dramatic effect on the specificity and strength of signal observed with an antibody (as is illustrated on the datasheet of https://www.abcam.com/ab9385). I would also suggest increasing the concentration of your SDS-page gel, from 8 % to 12 % to improve the resolution of the bands.

Should the suggestions not improve the results, please do let me know.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

Read More

Answer

Thank you for your kind feedback. I am glad to hear that the antibody finally works for you. I sincerely apologize for the inconvenience caused due to failing of lot GR50119-1.

We acknowledge your request and you being a valued customer of Abcam, we can consider providing discount. We have 28 vials of GR79241-1 in stock, could you tell me how many vials you would like to order?



I will look forward to hearing from you soon.

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1-10 of 27 Q&A

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