Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online

Anti-IDOL 抗体 (ab74562)

Reviews (4)Q&A (6)References (11)
Western blot - Anti-IDOL antibody (ab74562)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDOL antibody (ab74562)

Key features and details

  • Rabbit polyclonal to IDOL
  • Suitable for: ELISA, WB, IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-IDOL antibody

  • 製品の詳細

    Rabbit polyclonal to IDOL

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: ELISA, WB, IHC-Pmore details

  • 種交差性

    交差種: Mouse, Human

  • 免疫原

    Synthetic peptide corresponding to Human IDOL (internal sequence). Immunogen Range:161-210
    Database link: Q8WY64

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab74562の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
ELISA
Use at an assay dependent concentration.
WB (3)
1/500 - 1/1000. Predicted molecular weight: 50 kDa.
IHC-P (1)
1/50 - 1/100.
特記事項
ELISA
Use at an assay dependent concentration.
WB
1/500 - 1/1000. Predicted molecular weight: 50 kDa.
IHC-P
1/50 - 1/100.

ターゲット情報

  • 機能

    E3 ubiquitin-protein ligase that mediates ubiquitination and subsequent proteasomal degradation of myosin regulatory light chain (MRLC), LDLR, VLDLR and LRP8. Proteasomal degradation of MRLC leads to inhibit neurite outgrowth in presence of NGF by counteracting the stabilization of MRLC by saposin-like protein (CNPY2/MSAP) and reducing CNPY2-stimulated neurite outgrowth. Acts as a sterol-dependent inhibitor of cellular cholesterol uptake by mediating ubiquitination and subsequent degradation of LDLR.

  • 組織特異性

    Ubiquitously expressed.

  • パスウェイ

    Protein modification; protein ubiquitination.

  • 配列類似性

    Contains 1 FERM domain.
    Contains 1 RING-type zinc finger.

  • 発生段階

    Expressed in fetal tissues and higher levels were detected in placenta and fetal lung.

  • ドメイン

    The RING domain mediates ubiquitination and the neurite outgrowth inhibitory activity.

  • 翻訳後修飾

    Autoubiquitinated.

  • 細胞内局在

    Cytoplasm.
  • Information by UniProt

  • 参照データベース

    • 別名

      • BM-023 antibody
      • BZF1 antibody
      • E3 ubiquitin protein ligase MYLIP antibody
      • E3 ubiquitin-protein ligase MYLIP antibody
      • Idol antibody
      • Inducible degrader of the LDL-receptor antibody
      • Lipoprotein receptor antibody
      • MIR antibody
      • MYLIP antibody
      • MYLIP_HUMAN antibody
      • Myosin regulatory light chain interacting protein antibody
      • PP5242 antibody
      see all

    画像

    • Western blot - Anti-IDOL antibody (ab74562)
      Western blot - Anti-IDOL antibody (ab74562)
      All lanes : Anti-IDOL antibody (ab74562) at 1/500 dilution

      Lane 1 : extracts from HeLa cells
      Lane 2 : extracts from Jurkat cells
      Lane 3 : extracts from Jurkat cells with immunising peptide at 10 µg

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (H+L) HRP at 1/10000 dilution

      Predicted band size: 50 kDa
      Observed band size: 50 kDa
      Additional bands at: 35 kDa. We are unsure as to the identity of these extra bands.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDOL antibody (ab74562)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDOL antibody (ab74562)

      Immunohistochemistry analysis of IDOL in paraffin embedded human brain tissue using 1/50 ab74562.
      Left image: untreated sample.
      Right image: sample treated with immunising peptide.

    参考文献 (11)

    ab74562 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab74562 は 11 報の論文で使用されています。

    • Guo BC  et al. Di-(2-ethylhexyl) Phthalate Limits the Lipid-Lowering Effects of Simvastatin by Promoting Protein Degradation of Low-Density Lipoprotein Receptor: Role of PPARγ-PCSK9 and LXRα-IDOL Signaling Pathways. Antioxidants (Basel) 12:N/A (2023). PubMed: 36830035
    • Zhao L  et al. Long non-coding RNA TUSC8 inhibits breast cancer growth and metastasis via miR-190b-5p/MYLIP axis. Aging (Albany NY) 12:2974-2991 (2020). PubMed: 32039833
    • Ito S  et al. CNPY2 inhibits MYLIP-mediated AR protein degradation in prostate cancer cells. Oncotarget 9:17645-17655 (2018). PubMed: 29707137
    • Leitch EK  et al. Inhibition of low-density lipoprotein receptor degradation with a cyclic peptide that disrupts the homodimerization of IDOL E3 ubiquitin ligase. Chem Sci 9:5957-5966 (2018). PubMed: 30079210
    • Zhao L  et al. miR-19b promotes breast cancer metastasis through targeting MYLIP and its related cell adhesion molecules. Oncotarget 8:64330-64343 (2017). PubMed: 28969074
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 10 Abreviews or Q&A

    Western blot abreview for Anti-IDOL antibody

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MCF-7 (breast cancer))
    Gel Running Conditions
    Reduced Denaturing (8)
    Loading amount
    20 µg
    Specification
    MCF-7 (breast cancer)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 May 21 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-IDOL antibody

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Heart)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Sodium Citrate Buffer
    Permeabilization
    No
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Sep 01 2011

    Western blot abreview for Anti-IDOL antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Heart - left ventricle)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    50 µg
    Specification
    Heart - left ventricle
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Aug 30 2011

    Western blot abreview for Anti-IDOL antibody

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Heart - left ventricle)
    Gel Running Conditions
    Reduced Denaturing (8)
    Loading amount
    40 µg
    Specification
    Heart - left ventricle
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 May 27 2011

    Question

    Thank you once again for all your help!
    I have attached the completed form and an indicative WB for ab74562. The
    positive controls are at both ends. Could you send the replacement
    directly to me at the address below please?

    Read More
    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 123456.

    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.

    Read More

    Question

    WB shows no band, even for the pos control!

    Read More
    Answer

    Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    As we discussed on the phone, I am [ ATTACHING] our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

    I look forward to receiving your reply.

    Read More

    Question

    Actually markers were there, true it was not clear in the picture (there is a MW marker band with 50 next to it. I admit I did not make beautiful labels). I can confirm there was no band at 50kDa region (or unspecific band at 35kDa as suggested on Datasheet). I will test the one I have with other cell lysates before ask for a replacement.

    I have CHO lysate, J774 lysate, BHK lysate. Are those good + controls?

    Read More
    Answer

    Thank you for your reply.
    The CHO cells which are chinese hamster ovary cells as well as the BHK cells which are baby hamster kidney cells are untested species and I do not know if they would work with the antibody. Thus, these would not be suitable positive controls.
    But the J774 cells could be tried if they express the MYLIP protein.
    I look forward to hear back from you with your results.
    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Question

    I did not detect any signal (not even background, it was a clean film) first time with MYLIP ab, so abcam sent me another one as a replacement. With the new anti-MYLIP, I detect somethings, but there was not band (Please refer to the WB film picture). Do you have any suggestions?

    My Conditions:
    -Denatured gel (10% resolving, 4% stacking)
    -Tested on HepG2 cell lysate from another company (GAPDH confirmed the cell lysate was good)
    -50ug (20uL) proteins loaded, 5 min 95C treated.
    -Membrane transfer was verified by Ponceau Red Stain
    -5% skim milk blocking
    -primary antibody 1:500 in 1% skim milk ab-74562
    -secondary anti-rabbit antibody 1:5000 in 1% skim milk from another company
    -ECL reagent 5min
    -Exposure time 1h 15min

    I attached the WB film and the invoice.

    Read More
    Answer

    Thank you for contacting us.
    I am sorry to hear that this antibody is not giving you a band.
    The protocol you are using seems fine to me. The WB image you sent is out of focus and MW markers are not indicated, thus it is difficult to judge if the blot is completely empty or has any other faint bands.
    I would suggest using another lysate such as Hela or Jurkat as positive control.
    The protein seems to be expressed in fetal tissues and higher levels were detected in placenta and fetal lung. I am not sure about the expression levels in HepG2, though.
    I'd suggest trying either a different lysate where we know that MYLIP is expressed or trying another MYLIP antibody. I'd be happy to send you one or the other free of charge. Please let me know what approach you woud like to take.
    I look forward to hear back from you.

    Read More

    Question

    Lysis buffer: 20mM tris ph7.5 5mM EDTA 5mM or 5nM EGTA (i don't remember which one, but i used the right one) 0.5% maltosidase 1 tablet of mini protease inhibitor Wb conditions: 10% denaturing (with SDS) resolving gel, 4% stacking gel (with SDS) Samples were heated 95C 5 min I used denatured gel Ab concentration According to the quantity of protein i loaded and the exposure time, i don't think 1:500 will make a difference.

    Read More
    Answer

    Thank you for your message and for providing this further information.

    I appreciate the time you have spent on these experiments, and would like to offer a free of charge replacement for this product (providing the product has been purchased in the last six months). In order to arrange this, I would appreciate if you could confirm your order number and date of purchase?

    Thank you for your continued cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question

    I tested this antibody for Hamster hepatocytes (our sample 170ug protein loaded) and three controls (J774, CHO, HepG2); however, in neither of it I see a band. I attached the wb film with it, the film is very clean. Do you have any suggestions?   Here are the conditions I used: Our sample 170ug proteins, positive controls (J774, CHO, HepG2) more than 70 ug. The transfer was good, verified by Ponceau S stain 1:1 000 primary antibody in 1% milk TBS, room temperature 4 hours 1:5 000 secondary goat anti-rabbit antibody from Jackson Immunoresearch, Room temperature 2 hours Exposure time in the wb film I sent you is 1 hour, and it was right away after I drain the membrane.   I took the picture before I labeled the lanes. From left to right: My sample, J774, CHO, HepG2.  

    Read More
    Answer

    Thank you for contacting us.

    I am sorry to hear of the difficulties which you have been experiencing using this product. We do know that the control lysates which you have chosen should give result in western blot. Could you please tell me about the sample treatment for these cell lines, e.g. what type of lysis buffer you had used; Are these run under reduced/denatured conditions; Etc.?

    Have you attempted using a higher antibody concentration?

    This product is covered by our Abpromise guarantee. If we cannot remedy this issue and this is a product that you have purchased within the last six months, we will replace or refund it under our Abpromise guarantee, as you are using it according to specifications listed on our datasheet.
    I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.
    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com