Anti-ING4 抗体 (ab3714)

Submit a review Q&A (4)References (7)

Key features and details

  • Goat polyclonal to ING4
  • Suitable for: WB
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-ING4 antibody
    ING4 一次抗体 製品一覧

  • 製品の詳細

    Goat polyclonal to ING4

  • 由来種

    Goat

  • 特異性

    Tested on a U2OS cell line overexpressing human ING4.

  • アプリケーション

    適用あり: WBmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse, Rat, Dog

  • 免疫原

    Synthetic peptide:

    CLVRTSPEYGMPS

    , corresponding to amino acids 163 - 174 of Human ING4.

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

  • 特記事項


    ING4 is moderately similar to ING1 and may act in chromatin-mediated transcriptional regulation. ING4 contains a PHD-finger.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab3714 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB 1/500 - 1/3000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).

ターゲット情報

  • 機能

    Component of the HBO1 complex which has a histone H4-specific acetyltransferase activity, a reduced activity toward histone H3 and is responsible for the bulk of histone H4 acetylation in vivo. Through chromatin acetylation it may function in DNA replication. May inhibit tumor progression by modulating the transcriptional output of signaling pathways which regulate cell proliferation. Can suppress brain tumor angiogenesis through transcriptional repression of RELA/NFKB3 target genes when complexed with RELA. May also specifically suppress loss of contact inhibition elicited by activated oncogenes such as MYC. Represses hypoxia inducible factor's (HIF) activity by interacting with HIF prolyl hydroxylase 2 (EGLN1).

  • 配列類似性

    Belongs to the ING family.
    Contains 1 PHD-type zinc finger.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • Brain my036 protein antibody
    • Candidate tumor suppressor p33 ING 1 homolog antibody
    • Candidate tumor suppressor p33 ING1 homolog antibody
    • D6Wsu147e antibody
    • D6Xrf92 antibody
    • ING 1 like protein antibody
    • ING 4 antibody
    • ING1 like protein antibody
    • ING4 antibody
    • ING4_HUMAN antibody
    • Inhibitor of growth family member 4 antibody
    • Inhibitor of growth family member 4 long isoform antibody
    • Inhibitor of growth protein 4 antibody
    • MGC12557 antibody
    • my036 antibody
    • p29 ING 4 antibody
    • p29 ING4 antibody
    • p29ING4 antibody
    see all

画像

  • Western blot - Anti-ING4 antibody (ab3714)
    Western blot - Anti-ING4 antibody (ab3714)
    Anti-p29 ING4 polyclonal antibody detects ING4 protein by western blot. This antibody was used at 1.0 ug/ml to detect ING4 (lane 2) present in a U2OS whole cell lysate over expressing the protein. A control lysate (lane 3) shows no background staining. Comparison to MW markers (lane 1) indicates detection of a single band at ~29 kDa corresponding to ING4.

    A 4-20% Tris-glycine gradient gel was used to separate the protein by SDS-PAGE under reducing conditions. The protein was transferred to nitrocellulose using standard methods. After blocking using 5% non-fat dry milk in PBS, the membrane was probed with the primary antibody overnight at 4° C followed by washes and reaction with a 1:20,000 dilution of IRDye800 conjugated Rabbit anti Goat IgG [H&L] for 45 min at room temperature. LICOR's Odyssey® Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.

  • Western blot - Anti-ING4 antibody (ab3714)
    Western blot - Anti-ING4 antibody (ab3714)
    Anti-p29 ING4 polyclonal antibody detects ING4 protein by western blot in over expressed cell lysates. This antibody was used at 1.0 µg/ml to detect ING4 expression in control (-) and transformed U2OS and HeLa cell lysates. A predominant band corresponding to p29 ING4 is only seen in lysates from transformed cells. Anti-p29 ING4 polyclonal antibody detects ING4 protein by western blot in over expressed cell lysates. This antibody was used at 1.0 ug/ml to detect ING4 expression in control (-) and transformed U2OS and HeLa cell lysates. A predominant band corresponding to p29 ING4 is only seen in lysates from transformed cells.

参考文献 (7)

ab3714 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。

ab3714 は 7 報の論文で使用されています。

  • Culurgioni S  et al. Crystal structure of inhibitor of growth 4 (ING4) dimerization domain reveals functional organization of ING family of chromatin-binding proteins. J Biol Chem 287:10876-84 (2012). WB . PubMed: 22334692
  • Conner J & Braidwood L Expression of inhibitor of growth 4 by HSV1716 improves oncolytic potency and enhances efficacy. Cancer Gene Ther 19:499-507 (2012). WB . PubMed: 22595793
  • Moreno A  et al. Functional impact of cancer-associated mutations in the tumor suppressor protein ING4. Carcinogenesis 31:1932-8 (2010). WB ; Human . PubMed: 20705953
  • Li X  et al. Inhibitor of growth 4 induces growth suppression and apoptosis in glioma U87MG. Pathobiology 76:181-92 (2009). PubMed: 19571607
  • Li X  et al. ING4 induces cell growth inhibition in human lung adenocarcinoma A549 cells by means of Wnt-1/beta-catenin signaling pathway. Anat Rec (Hoboken) 291:593-600 (2008). PubMed: 18399550
  • Iizuka M  et al. Hbo1 Links p53-dependent stress signaling to DNA replication licensing. Mol Cell Biol 28:140-53 (2008). WB ; Human . PubMed: 17954561
  • Nozell S  et al. The ING4 tumor suppressor attenuates NF-kappaB activity at the promoters of target genes. Mol Cell Biol 28:6632-45 (2008). WB ; Human . PubMed: 18779315

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Question

DESCRIPTION OF THE PROBLEM wrong band size ,more bands, SAMPLE human tissure PRIMARY ANTIBODY ab3714 (1:1000;1:500;1:250)dilution:5%nonfat milk/TBS-t=1/4 incubation time:4`covernight ; room temperature 2h;room temperature 1h wash step:TBS-T 10min *2 TBS 10min *1 DETECTION METHOD ECL ANTIBODY STORAGE CONDITIONS subpackage and store -20 dilution store -4 SAMPLE PREPARATION 4*loading buffer ,15ulProtease inhibitors/ml.100`c 5-10min AMOUNT OF PROTEIN LOADED 60ug_100ug ELECTROPHORESIS/GEL CONDITIONS 15% TRANSFER AND BLOCKING CONDITIONS transfer buffer:tris/glycine 300ma 40min (the optimization time verified by ponceau red) blocking :5%nonfat milk 4`c overnight SECONDARY ANTIBODY HRP-antibody from rabbit (1:2000) incubation time(room temperature 2h;room temperature 1h;room temperature 40min) wash step:TBS-T 10min *2 TBS 10min *1 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 15 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES I have got no band when i use ab3714(1:3000) ab3714 lot:164571

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Answer

Thank you for your enquiry and for submitting your protocol details. I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionaire in association with your blot images and I would appreciate it if you could address the following questions; 1. Please can you confirm that the samples that you are running are tissue samples. 2. Can you tell me how you are preparing the tissue lysates; the details in the questionaire were confusing. What buffer are you using. 3. In an earlier email you mentioned that "The 29KD lane in the PVDF when is dealt with ponceau red". Does this meant that the marker lane in the blots that you emailed me has a molecular weight marker of 29KDa. 4. Can you tell me the percentage of Tween in the TBST that you are using. You mention "TBS-t=1/4". Can you please tell me the percentage of Tween 20 that you have been using. I ask because there is noticeable noise in the immunoblots that you have emailed me. 5. Can you tell me whether you have been able to obtain good results using this approach with an alternative antibody but the same tissue lysate. This would confirm that the approach, reagents, secondary antibody, transfer conditions etc that you are using are ok. I appreciate your patience and look forward to hearing from you.

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Question

We are trying both the immunohistochemistry and the western. While the immuno is promising and hopefully we will be able to come back with something soon; we are having some issues with the western. We tried the dilution of 1:1000 ING4 and 1:5000 secondary and did not get any bands. Can you give us the standardised method that your lab had used to confirm the use as Western including what secondary atibody is recommended along with the dilution. We did the loading control (tubulin) and the loading control looks allright. Will also be grateful if you can attach the western blot pictures for our ready reference.

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Answer

Thank you for your enquiry. I'm glad to hear that the antibody is giving good preliminary results in IHC. I hope that you will submit an Abreview with your results. Regarding the Western blot, the recommended starting dilution range is between 1/500 to 1/3000. It may be that you must use the antibody at a higher concentration (I generally recommend starting at 1/500 for polyclonals as well) to visualize the bands. It would be great if you could let me know further details of your protocol so we could see what kind of samples, how much protein, etc. The questionnaire for this is at the following link: https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=3714&mode=questionaire This antibody was used at 1.0 µg/ml to detect ING4 present in a U2OS whole cell lysate over expressing the protein. A single band was detected at ~29 kDa corresponding to ING4. A 4-20% TRISglycine gradient gel was used to separate the protein by SDS-PAGE under reducing conditions. The protein was transferred to nitrocellulose using standard methods. After blocking using 5% non-fat dry milk in PBS, the membrane was probed with the primary antibody overnight at 4° C followed by washes and reaction with a 1:20,000 dilution of IRDye800 conjugated Rabbit-anti-Goat IgG [H&L] for 45 min at room temperature. LICOR's Odyssey® Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results. The HRP-conjugated secondary recommended for this antibody is ab6741, Rabbit polyclonal to Goat IgG. I have added a new Western blot image to the datasheet (it may take a couple of hours for it to show up online). Unfortunately I am unable to attach the images to this email, but I have provided a link directly to the datasheet at the bottom of this email. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

BATCH NUMBER 63946 DESCRIPTION OF THE PROBLEM No any Band. SAMPLE Cell Lysate PRIMARY ANTIBODY ab3714 SECONDARY ANTIBODY ABcam/Rabbit anti Goat IgG HRP DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED Cell Lysate SAMPLE PREPARATION PBS Buffer TRANSFER AND BLOCKING CONDITIONS 0.5% milk in PBS HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 HAVE YOU RUN A "NO PRIMARY" CONTROL? No

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Answer

Thank you for your enquiry. We would like to emphasize the fact that this is a Fast track antibody and it has only been assayed by ELISA against 0.1µg of the immunizing peptide. It has not been tested for Western blot analysis at all. We have read through the completed questionnaire; however we still need the following information. Unfortunately, the details you have provided are not enough for us to be able to identify the source of the problem. What type of samples your customer has tested (species, cells, tissue, recombinant protein etc)? How was the cell lysate prepared? How much protein was loaded onto the gel? At what dilution the antibody was tested? Has the customer tested the detection system? How about the protein transfer? What did he/she use as positive control? We look forward to hearing from you soon.

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Question

Just a confirmation: my customer wants to use this ab in ELISA. I suggested him your ab6742 (Rabbit polyclonal to Goat IgG H&L) as a secondary antibody, because in the ELISA protocol you suggested to use an alkaline phosphatase marked ab. Did I give him the right suggestion? (today I have no technical support people in my office..)

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Answer

At the bottom of the online datasheet for ab3714 there is a list of compatible secondary antibodies and so yes, ab6742 is an appropriate choice.

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