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Anti-PML+RARA Fusion 抗体 (ab43152)

Reviews (1)Q&A (3)References (13)
Western blot - Anti-PML+RARA Fusion antibody (ab43152)

    Key features and details

    • Rabbit polyclonal to PML+RARA Fusion
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG

    リコンビナント抗体で、ロット間での高い再現性を実現

    Product image
    Anti-PML Protein antibody [EPR16792] (ab179466)
    • 異なるロット間での安定した再現性
    • 容易なスケールアップ
    • 評価試験による特異性の確認済み
    • 倫理基準に準拠 - アニマル・フリーの生産

    製品の概要

    • 製品名

      Anti-PML+RARA Fusion antibody

    • 製品の詳細

      Rabbit polyclonal to PML+RARA Fusion

    • 由来種

      Rabbit

    • アプリケーション

      適用あり: WBmore details

    • 種交差性

      交差種: Human

    • 免疫原

      Synthetic peptide corresponding to Human PML+RARA Fusion aa 500-600 conjugated to keyhole limpet haemocyanin.
      (Peptide available as ab46898)

    • 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ab43152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
    WB (1)
    Use at an assay dependent concentration. Predicted molecular weight: 98 kDa.
    特記事項
    WB
    Use at an assay dependent concentration. Predicted molecular weight: 98 kDa.

    ターゲット情報

    • 関連性

      PML protein is a member of the tripartite motif (TRIM) family. The TRIM motif includes three zinc-binding domains, a RING, a B-box type 1 and a B-box type 2, and a coiled-coil region. This phosphoprotein localizes to nuclear bodies where it functions as a transcription factor and tumor suppressor. Its expression is cell-cycle related and it regulates the p53 response to oncogenic signals. Retinoic acid, a metabolite of vitamin A, is necessary for normal organogenesis but acts as a teratogen at high levels during embryonic and fetal development. Retinoic acid functions through its interaction with the nuclear protein, retinoic acid receptor (RAR). RAR belongs to the steroid and thyroid hormone superfamily of nuclear receptor proteins which exert their effects by binding to specific DNA response elements, thus regulating gene expression in target cells. RAR exists as three major subtypes: alpha, beta and gamma.

    • 細胞内局在

      Nuclear and Cytoplasmic

    • 参照データベース

      • 別名

        • MYL antibody
        • NR1B1 antibody
        • Nuclear receptor subfamily 1 group B member 1 antibody
        • Nucleophosmin-retinoic acid receptor alpha fusion protein NPM-RAR long form antibody
        • PP8675 antibody
        • Promyelocytic leukemia antibody
        • Promyelocytic leukemia protein antibody
        • Promyelocytic leukemia, inducer of antibody
        • RAR antibody
        • RAR-alpha antibody
        • RARA alpha antibody
        • RARAalpha1 antibody
        • Retinoic acid receptor alpha polypeptide antibody
        • Retinoic acid receptor, alpha antibody
        • RING finger protein 71 antibody
        • RNF71 antibody
        • TRIM19 antibody
        • Tripartite motif protein TRIM19 antibody
        • Tripartite motif-containing protein 19 antibody
        see all

      画像

      • Western blot - Anti-PML+RARA Fusion antibody (ab43152)
        Western blot - Anti-PML+RARA Fusion antibody (ab43152)Image courtesy of Christian Marx (University of Jena, Germany)

        Anti-PML+ RARα Fusion antibody (ab43152) reactivity with reduced NB4 (A and B) and K562 (B) cell lysate. After SDS-PAGE (where 100μg lysate was loaded in A and 20μg lysate in B), membranes were blocked in 5% Milk in PBS + 0.1% Tween for 1h at 25°C before incubation with ab43152 (1:5,000 dilution in 1% Milk PBS + 0.1% Tween) for 16h at 4ºC. Blots was then incubated with an anti-Rabbit HRP-conjugated secondary antibody before developing with ECL.

      プロトコール

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      参考文献 (13)

      ab43152 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。

      ab43152 は 13 報の論文で使用されています。

      • Villiers W  et al. Multi-omics and machine learning reveal context-specific gene regulatory activities of PML::RARA in acute promyelocytic leukemia. Nat Commun 14:724 (2023). PubMed: 36759620
      • Qi X  et al. HDN-1 induces cell differentiation toward apoptosis in promyelocytic leukemia cells depending on its selective effect on client proteins of Hsp90. Toxicol Appl Pharmacol 417:115459 (2021). PubMed: 33609515
      • Zhao M  et al. OGP46 Induces Differentiation of Acute Myeloid Leukemia Cells via Different Optimal Signaling Pathways. Front Cell Dev Biol 9:652972 (2021). PubMed: 33748146
      • Della Via FI  et al. (-)-Epigallocatechin-3-gallate induces apoptosis and differentiation in leukaemia by targeting reactive oxygen species and PIN1. Sci Rep 11:9103 (2021). PubMed: 33907248
      • Li S  et al. A novel all-trans retinoic acid derivative regulates cell cycle and differentiation of myelodysplastic syndrome cells by USO1. Eur J Pharmacol 906:174199 (2021). PubMed: 34058203
      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

      1-4 of 4 Abreviews or Q&A

      Western blot abreview for Anti-PML+RARA Fusion antibody

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Loading amount
      100 µg
      Gel Running Conditions
      Reduced Denaturing (8 %)
      Sample
      Human Cell lysate - whole cell (NB4 and K562 human cell lines)
      Specification
      NB4 and K562 human cell lines
      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.

      MR. Christian Marx

      Verified customer

      投稿 Jul 31 2013

      Question

      Dear Tech Support Support Team,



      Our customer is interested in ab43152. Kindly advise the positive control for this antibody in WB. The customer will perform WB with human samples. Will the customer get Abpromise for WB, human samples if this application is shown in the reference article? Will it be possible to proceed with the Abreview promotion in the case that the Abpromise is not acceptable by you?



      Thanks in advance for your reply.




      Have a nice week,

      Read More
      Answer

      Thank you for enquiry and your interest.
      This product is Fast track and as these antibodies have not been fully characterized at Abcam, we cannot guarantee that the antibody will work in any application other than ELISA against the immunizing peptide and they are therefore not covered by the full Abpromise guarantee. We sell at a lower price and can award much higher Abpoints for reviewing (950 - see more details on the datasheet).
      Positive control can be human acute promyelocytic cell such as HL60 or NB4 cell lines.
      I hope this helps and if I can assist further, please do not hesitate to contact me.

      Read More

      Question

      Bonjour, Nous avons récemment utilisé un anticorps Abcam anti-PML-RAR (ab 43152, lot 653110) qui nous donnait toute satisfaction. Il nous a fallu renouveler cet anticorps. Malheureusement le nouvel anticorps reçu (lot 885950) ne permet pas la visualisation de la protéine PML-RAR. Nous avons confirmé que le problème venait bien de l'anticorps en réhybridant la même membrane avec un aliquot qui nous restait du précédent lot. Je vous joins une image des blots obtenus après hybridation avec l'anti-PML+RAR (ab 43152, 1/1 000). L'hybridation 1 a été faite avec ab 43152, lot 885950. L'hybridation 2 a été faite avec ab 43152, lot 653110 sur la même membrane. Nous en concluons que le lot 885950 ne permet pas de détecter la protéine PML-RAR. Ce résultat a été confirmé deux fois. Avez vous eu des retours concernant ce lot d'anticorps? Pouvez vous nous proposez un lot de remplacement? Dans l'attente de votre réponse, Bien cordialement

      Read More
      Answer

      Merci de nous avoir contactés. Nous sommes désolés d'apprendre que le produit que vous avez reçu ne fonctionne pas comme attendu. Nous allons vous remplacer volontiers ce tube fautif avec un nouveau tube. Veuillez me confirmer pour ceci votre numéro et date de commande, ainsi que si vous souhaitez accepter cette offre. De plus, je vous serait reconnaissante si vous pourriez me confirmer sur quel type et espèce de tissue vous avez testé ces anticorps. Nous avons effectivement reçu un autre retour négative sur ce lot. Nous investiguons tous les retours négatives et allons aussi essayer de trouver ce qui s'est passé avec ce lot. La qualité de nos produits et très important pour nous. Je suis très désolée pour ce tube fautif. Je me réjouis de votre réponse.

      Read More

      Question

      Dear Technical Team, Our customer purchased Ab43152. They tried Western Blot using this anibody but did not have a good luck to get a data. They want to ask your opinion how to imporve their experimental procdure. Could you please review their data and let us know how you think? (Please check the files I attached) FYI, they chose this antibody because of the reference shown on your website. When he bought this Item, he strongly believed that it would work because Reference was shown on the website. They could not understand why it worked well in the authors of the paper but it did not work in their hands. Thank you in advance for your help.    Best regards,     Research Purpose I’m going to introduce briefly what we do. We had been studying on differentiation induction and its blocker PML-RARA in model of Acute Promylocytic Leukemia. Also we’re focusing role of PML-RARA degradation via various mechanism. Correlation of these is well known.     1) Abcam product code  ab                     Ab43152 2) Abcam order reference number or product batch number                     885950 2-1) storage temperature of antibody After receiving that, I aliquot and then stored at -70 ℃ in defreezer. 3) Description of the problem Your product could not detect PML-RARA, which is target band 4) Sample preparation: NB4, which is leukemia cell line Lysis buffer : RIPA buffer Protease inhibitors: complete product from Roche(cat # 11873580001) Phosphatase inhibitors : phosphatase inhibitor cocktail I & II from Sigma(cat# P2850 & P5726) Reducing agent beta-mecaptoethanol Boiling for ≥5 min Protein loaded ug/lane or cells/lane: I loaded 40 ug protein of NB4, which is leukemia cell line involving endogenous PML-RARa gene Positive control PML-RARa transfected 293T lysate Negative control : 293T Wild type lysate 5) Percentage of gel Type of membrane : nitrocellulose membrane Protein transfer verified: using Ponceau S staining solution Blocking agent and concentration: 5% non-fat milk Blocking time : 1h  Blocking temperature : Room Temperature 6) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution :  1:500 Diluent buffer  1X TES-T buffer in shaking Incubation time : for 14h in shaking Incubation temperature:  4℃ in refrigerator 7)  Secondary antibody: Species:goat Isotype: IgG Reacts against: anti-rabbit Concentration or dilution : 1:500 Diluent buffer : 5% non-fat milk Incubation time : 1h Incubation temperature: at room temperature Fluorochrome or enzyme conjugate: 8) Washing after primary and secondary antibodies: Buffer :  1X TES-T Number of washes : 4 times for 10min, respectively 9) Detection method ECL 10) How many times have you run this staining? Exactly 2 times, I have - Do you obtain the same results every time?  Absolutely, yes - Have you run a "No Primary" control? Not this time, but I had it, so that I did not any problem that you ask - What steps have you altered to try and optimize the use of this antibody? I tried to dilute primary antibody condition from 1:1000 to 1:500 Please check the attached image    

      Read More
      Answer

      Thank you for taking the time to complete our questionnaire and contact us. 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 this is also helpful in our records for monitoring of quality. Reviewing this case, I would like to offer some suggestions to help optimize the results from ab43152. I would also appreciate if you can confirm some further details: 1. Please confirm the Abcam order reference number. Was this 974371 (PO1199)? 2. I am sorry to confirm that this is a Fast track antibody. This means it has not been well characterized yet, and so we sell it as a lower price. We are not able to provide our guarantee for Fast Track antibodies. However, the customer may like to consider submitting an  Abreview, as for a Fast Track antibody, this is worth 900 Abpoints.  Further details and terms and conditions regarding our Fast Track antibodies can be found on the Fast Track link on the datasheet. I have attached this information as a document. 3. Could you confirm exactly what is in each lane of the image: PML-RARa lane:  is this standard purified PML-RARa protein? Wild type:  Is this just cells with no treatment? 1 um ATRA cells that have been treated? 4. The NB4 cells are showing a nice band at 70 kDa. According to SwissProt/Uniprot there are several isoforms of the protein that have a molecular weight of about 70 kDa:   http://www.uniprot.org/uniprot/P29590 In fact there are a total of 14 isoforms of this protein. it may be the two different antibodies are detecting different isoforms, at different molecular weights. And it may be that the 293 cells do not express an isoform detected by ab43152. 4. I can recommend to try BSA rather than milk to block. Changing blocking agent can sometimes improve results. 5. I can recommend to  ensure cells are not overconfluent (70-80%), and to check the viability (which should be >95%). 6. I can suggest to try  fresh lysate, particularly for the 293T and to check again the protein concentration in each lysate. 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.  

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