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Anti-HDAC4 抗体 (ab16339)

Submit a review Q&A (6)References (1)
Western blot - Anti-HDAC4 antibody (ab16339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC4 antibody (ab16339)

Key features and details

  • Rabbit polyclonal to HDAC4
  • Suitable for: IHC-P, WB
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-HDAC4 antibody
    HDAC4 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to HDAC4

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IHC-P, WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide corresponding to Human HDAC4 aa 1-18.
    Sequence:

    MSSQSHPDGLSGRDQPVE

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • WB: HeLa cell lysate. IHC-P: Human skin tissue.

  • 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ab16339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
IHC-P
1/20.
WB
1/1000. Predicted molecular weight: 140 kDa.
特記事項
IHC-P
1/20.
WB
1/1000. Predicted molecular weight: 140 kDa.

ターゲット情報

  • 機能

    Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Involved in muscle maturation via its interaction with the myocyte enhancer factors such as MEF2A, MEF2C and MEF2D.

  • 組織特異性

    Ubiquitous.

  • 関連疾患

    Defects in HDAC4 are the cause of brachydactyly-mental retardation syndrome (BDMR) [MIM:600430]. A syndrome resembling the physical anomalies found in Albright hereditary osteodystrophy. Common features are mild facial dysmorphism, congenital heart defects, distinct brachydactyly type E, mental retardation, developmental delay, seizures, autism spectrum disorder, and stocky build. Soft tissue ossification is absent, and there are no abnormalities in parathyroid hormone or calcium metabolism.

  • 配列類似性

    Belongs to the histone deacetylase family. HD type 2 subfamily.

  • ドメイン

    The nuclear export sequence mediates the shuttling between the nucleus and the cytoplasm.

  • 翻訳後修飾

    Phosphorylated by CaMK4 at Ser-246, Ser-467 and Ser-632. Phosphorylation at other residues is required for the interaction with 14-3-3.
    Sumoylation on Lys-559 is promoted by the E3 SUMO-protein ligase RANBP2, and prevented by phosphorylation by CaMK4.

  • 細胞内局在

    Nucleus. Cytoplasm. Shuttles between the nucleus and the cytoplasm. Upon muscle cells differentiation, it accumulates in the nuclei of myotubes, suggesting a positive role of nuclear HDAC4 in muscle differentiation. The export to cytoplasm depends on the interaction with a 14-3-3 chaperone protein and is due to its phosphorylation at Ser-246, Ser-467 and Ser-632 by CaMK4. The nuclear localization probably depends on sumoylation.
  • Information by UniProt

  • 参照データベース

    • 別名

      • AHO3 antibody
      • BDMR antibody
      • EC 3.5.1.98 antibody
      • HA6116 antibody
      • HD 4 antibody
      • HD4 antibody
      • HDAC 4 antibody
      • HDAC A antibody
      • HDAC4 antibody
      • HDAC4_HUMAN antibody
      • HDACA antibody
      • Histone deacetylase 4 antibody
      • Histone Deacetylase A antibody
      • KIAA0288 antibody
      see all

    画像

    • Western blot - Anti-HDAC4 antibody (ab16339)
      Western blot - Anti-HDAC4 antibody (ab16339)
      ab16339 at 1/1000 detecting HDAC4 from HeLa cell lysate by Western blot
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC4 antibody (ab16339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC4 antibody (ab16339)

      Immunohistochemistry was performed on normal biopsies of deparaffinized Human skin tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a Rabbit Polyclonal Antibody recognizing HDAC4 (ab16339 ) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    参考文献 (1)

    ab16339 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。

    ab16339 は 1 報の論文で使用されています。

    • Shi L  et al. LncRNA MALAT1 promotes decidualization of endometrial stromal cells via sponging miR-498-3p and targeting histone deacetylase 4. Cell Biol Int 46:1264-1274 (2022). PubMed: 35616349

    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

    1-6 of 6 Abreviews or Q&A

    Question

    You say that " the immunogen sequence is KRPDEEPMEEEPPL, corresponding to amino acids 1-18 of Human HDAC4, not C-terminal as previously stated". I would like to clarify this because, as far as I am aware, the KRPDEEPMEEEPPL epitope is C-terminal and does not correspond to aa 1-18 of the HDAC4 protein. Could you please clarify this?

    Read More
    Answer

    Thank you for contacting us.

    I wish to apologize for the confusion as I had made this mistake when creating the recall email. The correct immunogen does correspond to aa1-18 of human HDAC4 and not to the peptide stated above. Thank you very much for alerting me to this mistake.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Question

    Dear xxxx,

    Just to let you know that the C-terminal antibody was all OK in that it recognised the full-length HDAC4 along with my C-terminal (but not N-terminal) constructs. Therefore I will now start using ab32073 instead of ab16339.

    Many Thanks,

    Read More
    Answer

    Thank you very much for sharing the results you obtained with ab32073. I am very pleased to hear that it has worked well for you.

    It would be very useful for other customers if you could share this information in the form of an Abreview? This should only take 5-10 minutes to complete and will allow other customers to judge if the antibody is appropriate for their experiments. As a reward you will earn Abpoints which can be converted to Amazon vouchers, or credit off future orders with Abcam.

    More information on the Abreview system and Abpoints can be found from the following link:

    https://www.abcam.com/abreviews

    https://www.abcam.com/abpoints

    If I can be of any further assistance, please do let me know.

    Until then, I wish you all the best with your research.

    Read More

    Question

    Dear xxxx
    No worries. OK, well at least that has resolved that issue and saved me running more gels and experiments today.

    I will process the xxx blots today, as they have been run, for completion.

    In terms of ab32073, yes if you send it over I can try it out for you and let you know if it works either way. I still don’t have a C-terminal antibody to recognise these constructs, so I could use this if all works OK. Hopefully the 2 amino acids difference will not make a difference. What is the exact epitope? The datasheet just said “C-terminal to HDAC4”.

    Best Regards,

    Read More
    Answer

    Thank you for your reply and understanding. I emailed as quickly as I could after receiving the information so that you wouldn't be running blots unnecessarily.

    I have now arranged for the rabbit monoclonal ab32073 to be sent to you. This is on the order number 1179028 (Purchase order number FOCR 4507028970/50A PRU2910995-1). This should reach you tomorrow.



    Please do let me know how the ab32073 performs in your experiments.

    In the meantime, if I can be of any further assistance, please do not hesitate to let me know.

    Read More

    Question

    Hi,

    OK, I will re-run the samples at some point tomorrow/Wednesday.

    With regards to aliquoting, no this was the first time I used it so it was (am still is) in the original abcam tube you sent. Also, because of this ab16339 was at -20 storage as it had been unused, whilst xxx was at +4oC as it had been opened. Therefore this is why I am am absolutely sure I have not mixed anything up as it is pretty much impossible.

    I will have the xx result tomorrow, so will let you know.

    If this the xxx result is as expected, I don’t know whether it will be worth sending ab32073 anyway? Up to you? But if I had that antibody here I could essentially map your ab1339 and ab32073 in the same experiment? Mind, the only problem I may see with ab32073 is that your website states it does not cross-react with mouse HDAC4 and these constructs I made are the mouse HDAC4 gene. But, who knows, may be in an over-expression system they will work?

    I’ll let you know tomorrow how I am getting on.

    Thanks,

    Read More
    Answer

    Thank you for getting back to me.

    As I think I mentioned to you, I was consulting with the source of this antibody to get confirmation of the immunogen used to raise the antibody.

    Unfortunately, they have gone back in their records and found that an error was made. As your results have supported, the antibody was actually raised against the N-terminal portion of the HDAC4 protein. I am very sorry for this error and the inconvenience it has caused you. We will be updating our datasheet with this information, as well as emailing all former customers of this updated information. Thank you for bringing this to our attention.

    I will of course be able to send you the rabbit monoclonal ab32073 if you would like to try it. I am sorry but when I suggested it I was under the impression that you were using human sequences for your construct. You are correct that we have found this antibody not to be able to detect the mouse protein. This is as you have mentioned, with the endogenously expressed protein and with the over expressed constructs you are using this may be different. The immunogen used to raise the antibody has two alterations between the human and mouse sequence out of a total of 17 residues. We would therefore normally predict that there would be cross reactivity however I cannot guarantee this. If you find this antibody not to work in your experiments I would of course be able to provide you with a refund or credit note.

    If you would like me to send ab32073 please do let me know. If you let me know before 3 pm today it can be with you tomorrow.

    Alternatively, I can offer a credit note or refund.

    I am very sorry for the inconvenience this error has caused you. look forward to hearing how you would like to proceed.

    Read More

    Question

    Dear xxxxx,

    Thanks for your reply.

    Yes both antibodies are indeed anti-rabbit and therefore the non-specific profile (not surprisingly) would be the same. It is, however, exactly the same samples loaded onto duplicate gels and I probed the resultant blots separately in sealed plastic bags, therefore there is no way one antibody could have spilled into another. Also, I never combine membranes (so to speak) after the primary probe, even if they do have the same secondary. Therefore I am 99% sure I have not made an error.

    With regards to the xxxxxx antibody, that is exactly the experiment I am running today, and I will have this result tomorrow. Therefore hopefully this antibody will recognise the full-length protein, all N-terminal fragments, but not some of the more C-terminal fragments.

    If you like, to be absolutely 100% sure, I could re-run the samples and just probe them with ab16339 again today, then we would know for sure if it is an N-terminal antibody?

    Meanwhile, the rabbit ab32073 would be a great alternative for me to try also. The purchase order requisition number you mention is correct for this order.

    With best regards,

    Read More
    Answer

    Thank you for getting back to me. If you wouldn't mind testing ab16339 again I think that would be worthwhile, just to confirm there was no mix up along the way. If you get the same results again I'd be happy to send ab32073 if you would like to try it.

    I was just wondering, there is no change when aliquotting ab16339 to be frozen that any of the aliquots got mixed up with the other antibody?

    I'd be interested to see the results of both ab16339 and the xxxxx antibody. Hopefully the results are as would be expected this time.

    I look forward to receiving your reply.

    Read More

    Question

    Dear xxxxx,

    Further to our telephone conversation, please find a powerpoint presentation for HDAC4 consisting of three slides.

    Slide1, shows the constructs I have generated ranging from the full-length HDAC4 protein (H4-1) and various N-terminal (H4-2, H4-6 to H4-11) or C-terminal (H4-3 to H4-5) deletion mutants.

    Slide 2, shows them in vitro translated using a rabbit reticulolysate system.

    Slide 3, shows a HEK293T transfection with these constructs in induced or un-induced cells which over-express mutant Huntingtin (i.e. – or +), the disease in which we work on. As you will see I have probed blots with either the Sigma antibody DM-15 (top panels) or your abcam antibody ab16339 (bottom panels), and basically both gel sets look pretty much identical which suggests to me that the ab16339 clone you sell has to be N-terminal too. Therefore, as you will see there is no signal in mock untransfected cells (as expected) with either antibody. Both antibodies (as expected) recognise cells transfected with the full-length protein (H4-1). Then Sigxxxx(aa14-28) as expected should only recognise N-terminal fragments, i.e. H4-3, H4-4, and H4-5, whereas ab16339 (aa1071-1084) should only recognise H4-2, H4-6, H4-8, H4-9, H4-10 and H4-11.

    This, however, is not the case. In fact ab16339 recognises exactly the same as xxx implying that its epitope must indeed by N-terminal. As explained, there is no chance I have put the wrong antibodies on the wrong blots as the background (non-specific) banding profile is different between the xxx and ab16339 probed blots.

    Please let me know if you require any further information regarding this matter.

    With best regards,

    Read More
    Answer

    Thank you for providing that information. It has made understanding the problems encountered in your experiment much easier to understand.

    As we discussed over the phone, it is very unlikely that you have been provided with the wrong vial of antibody. From the results you have shared with me, I think the two most likely scenarios is that either the incorrect antibody was loaded on the membrane or that the antibody immunogen information is incorrect. I am currently looking into the latter.

    Although you have said that you are sure that the correct antibody was loaded the non-specific staining between the two antibodies does look very similar, with just a difference in intensity of the bands. This does suggest that it may be the same antibody. Can I assume both are raised in rabbit and the secondary antibody would therefore not be able to distinguish between the N-terminal antibody and ab16339?

    You mention another antibody, SC-H4, how did this perform in your experiment?

    In order to try to ascertain which reason is likely to be responsible for these unexpected results I can provide you with a different antibody to try free of charge. The rabbit monoclonal https://www.abcam.com/HDAC4-antibody-Y417-ab32073.html which is also raised against the C-terminal domain of the HDAC4.

    If you would like me to arrange this for you please do let me know. I currently have the order number xxxx (Purchase order number xxxxx) of the 5th of September 2012, is this correct?

    I look forward to receiving your reply.

    Read More

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