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Anti-CEBP Alpha/CEBPA 抗体 (ab15048)

Reviews (2)Q&A (5)References (10)
Western blot - Anti-CEBP Alpha/CEBPA antibody (ab15048)

    Key features and details

    • Rabbit polyclonal to CEBP Alpha/CEBPA
    • Suitable for: WB
    • Reacts with: Mouse
    • 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-CEBP Alpha/CEBPA antibody [EPR27924-90] (ab317442)
    • 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-CEBP Alpha/CEBPA antibody
      CEBP Alpha/CEBPA 一次抗体 製品一覧

    • 製品の詳細

      Rabbit polyclonal to CEBP Alpha/CEBPA

    • 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: Mouse

    • 免疫原

      Recombinant fragment corresponding to Rat CEBP Alpha/CEBPA.
      Database link: NM_012524

    • 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ab15048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 (2)
    Use a concentration of 2 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 30,43 kDa).

    Detects bands of approximately 30 kDa and 43 kDa representing CEBP alpha in mouse 3T3-L1 cells.
    特記事項
    WB
    Use a concentration of 2 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 30,43 kDa).

    Detects bands of approximately 30 kDa and 43 kDa representing CEBP alpha in mouse 3T3-L1 cells.

    ターゲット情報

    • 機能

      C/EBP is a DNA-binding protein that recognizes two different motifs: the CCAAT homology common to many promoters and the enhanced core homology common to many enhancers.

    • 配列類似性

      Belongs to the bZIP family. C/EBP subfamily.
      Contains 1 bZIP domain.

    • 細胞内局在

      Nucleus.
    • Information by UniProt

    • 参照データベース

      • 別名

        • Apoptotic cysteine protease antibody
        • Apoptotic protease Mch 5 antibody
        • C/EBP alpha antibody
        • C/ebpalpha antibody
        • CAP4 antibody
        • Caspase 8 precursor antibody
        • CBF-A antibody
        • CCAAT Enhancer Binding Protein alpha antibody
        • CCAAT/enhancer binding protein (C/EBP), alpha antibody
        • CCAAT/enhancer-binding protein alpha antibody
        • CEBP antibody
        • CEBP A antibody
        • CEBP alpha antibody
        • Cebpa antibody
        • CEBPA_HUMAN antibody
        • FADD homologous ICE/CED 3 like protease antibody
        • FADD like ICE antibody
        • FLICE antibody
        • ICE like apoptotic protease 5 antibody
        • ICE8 antibody
        • MACH antibody
        • MCH5 antibody
        • MORT1 associated CED 3 homolog antibody
        see all

      画像

      • Western blot - Anti-CEBP Alpha/CEBPA antibody (ab15048)
        Western blot - Anti-CEBP Alpha/CEBPA antibody (ab15048)
        Anti-CEBP Alpha/CEBPA antibody (ab15048) + 3T3-L1 cell lysate.

        Predicted band size: 30,43 kDa



        Western blot of CEBP alpha using ab15048 on 3T3-L1 cells.

      参考文献 (10)

      ab15048 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。

      ab15048 は 10 報の論文で使用されています。

      • Lee SG  et al. TGFBI remodels adipose metabolism by regulating the Notch-1 signaling pathway. Exp Mol Med 55:520-531 (2023). PubMed: 36854775
      • Venigalla S  et al. MED12 Regulates Human Adipose-Derived Stem Cell Adipogenesis and Mediator Kinase Subunit Expression in Murine Adipose Depots. Stem Cells Dev 31:119-131 (2022). PubMed: 35018809
      • Huang Y  et al. Small-Molecule Targeting of Oncogenic FTO Demethylase in Acute Myeloid Leukemia. Cancer Cell 35:677-691.e10 (2019). PubMed: 30991027
      • Fatima LA  et al. Estradiol stimulates adipogenesis and Slc2a4/GLUT4 expression via ESR1-mediated activation of CEBPA. Mol Cell Endocrinol 498:110447 (2019). PubMed: 31100494
      • Jung M  et al. Inhibiting casein kinase 2 overcomes paclitaxel resistance in gastric cancer. Gastric Cancer 22:1153-1163 (2019). PubMed: 31098863
      • Lee D  et al. HMGB2 is a novel adipogenic factor that regulates ectopic fat infiltration in skeletal muscles. Sci Rep 8:9601 (2018). PubMed: 29942000
      • Jang MK  et al. Histone H3K9 Demethylase JMJD2B Activates Adipogenesis by Regulating H3K9 Methylation on PPAR? and C/EBPa during Adipogenesis. PLoS One 12:e0168185 (2017). WB . PubMed: 28060835
      • Ma Y  et al. Highly efficient genome editing of human hematopoietic stem cells via a nano-silicon-blade delivery approach. Integr Biol (Camb) 9:548-554 (2017). PubMed: 28513735
      • Ptasinska A  et al. Identification of a dynamic core transcriptional network in t(8;21) AML that regulates differentiation block and self-renewal. Cell Rep 8:1974-1988 (2014). WB . PubMed: 25242324
      • Khanna-Gupta A  et al. Up-regulation of Translation Eukaryotic Initiation Factor 4E in Nucleophosmin 1 Haploinsufficient Cells Results in Changes in CCAAT Enhancer-binding Protein a Activity: IMPLICATIONS IN MYELODYSPLASTIC SYNDROME AND ACUTE MYELOID LEUKEMIA. J Biol Chem 287:32728-37 (2012). WB . PubMed: 22851180

      レビューと Q&A

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      1-7 of 7 Abreviews or Q&A

      Western blot abreview for Anti-CEBP Alpha/CEBPA antibody

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Cell lysate - other (differentiated adipocyte)
      Gel Running Conditions
      Non-reduced Denaturing (12.5%)
      Loading amount
      20 µg
      Specification
      differentiated adipocyte
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°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 Jan 21 2014

      Western blot abreview for Anti-CEBP Alpha/CEBPA antibody

       Good
      Abreviews
      Application
      Western blot
      Sample
      Mouse Cell lysate - whole cell (White andipose tissue)
      Specification
      White andipose tissue
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
      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 Oct 31 2005

      Question

      Is the blocking peptide for this antibody available?

      Read More
      Answer

      Thank you for contacting us. Unfortunately the blocking peptide is not available for purchase.

      Please do not hesitate to contact us if you need any more advice or information.

      Read More

      Question

      Our customer had some question about ab15048 antibody. 1. The immungen which you mentioned is NM_012524 that could be wrong. Becouse of the seqence is 1389 bp, it could't be recognized by ab15048 on 466-831 amino acid. http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=81295399 2.They encountered some trouble in WB that the data showed many non-specific bands. Could you please check these conditions below? 1. Order details: a.. Batch number: b.. Abcam order or Purchase order number: ab15048 c.. Antibody storage conditions (temperature/reconstitution etc) -20? 2. Please describe the problem (high background, wrong band size, more bands, no band etc). Many non-specific bands and I don’t know which band is target. 3. On what material are you testing the antibody in WB? · Species: mouse NIH/3T3 cell and 3T3-L1 cell · Cell extract or Nuclear extract: cell extract · Purified protein or Recombinant protein: neither, total cell extract as sample 3. The lysate a.. How much protein was loaded: 35 ug per lane b.. What lysis buffer was used: RIPA c.. What protease inhibitors were used: cocktail (pancreas extract, pronase, thermolysin, chymotrypsin, trypsin, papain) d.. What loading buffer was used: 5X sample buffer (625mM Tris-HCl pH6.8, 25% glycerol, 2% SDS, Bromopheol blue, 5% 2-ME) e.. Did you heat the samples: temperature and time: 97 ? 10min 4. Electrophoresis/Gel conditions/ Transfer conditions a.. Reducing or non reducing gel: Reducing b.. Gel percentage : 12% c.. Transfer conditions: 100V 4? 1hr 5. Blocking conditions a.. Buffer: PBST (0.1% Tween 20 in PBS) b.. Blocking agent: milk, BSA, serum, what percentage: 5% milk c.. Incubation time: 4? O/N d.. Incubation temperature: RT 6. Primary Antibody a.. Specification (in which species was it raised against): rabbit polyclonal Ab b.. At what dilution(s) have you tested this antibody: 1/1000 c.. What dilution buffer was used: 5% milk in PBST d.. Incubation time: O/N e.. Incubation temperature: 4? f.. What washing steps were done: 3 times each for 10 min wash with PBST 7. Secondary Antibody a.. Specification (in which species was it raised against)? Goat anti- rabbit IgG conjugated HRP b.. At what dilution(s) have you tested this antibody: 1/5000 c.. Incubation time: 1hr d.. Wash steps: 3 times each for 10 min with PBST e.. Do you know whether the problems you are experiencing come from the secondary? No 8. Detection method ECl, ECl+, other detection method: ECl 9. Background bands · Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): no · Is the blocking step sufficient? I have tested 4? O/N Yes · Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) I have washed 4 times and no difference · At what size are the bands migrating? No, because there are many bands and it’s not migrating bands. · Could they be degradation products of your target? There are non-specific bands at large size. · Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) 1 n 2p 3n 4p 11. Did you apply positive and negative controls along with the samples? Please specify. Yes, lane 1 and 3 are negative and lane 2 and 4 are positive controls 10. Optimization attempts · How many times have you tried the Western? 5 times · Do you obtain the same results every time e.g. are background bands always in the same place? YES · What steps have you altered? First, Prolong blocking time, 4? O/N Second, dilute the primary Ab with PBST or 5% skim milk in PBST. Third, take the liver and fat as positive controls but there are no target bands at all.

      Read More
      Answer

      I have received the following feedback from the originator of ab15048: According to the original researcher, the "Rabbit anti-C/EBP alpha antibody was created against c/EBP alpha recombinant protein, derived from nucleotide sequence 466-813 of rat origin (NM_012524)" This antibody should detect bands at ~30 and ~43 kDa, which your customer is detecting among the other, non-specific bands. According to the original researcher, "HeLa cells (80% confluent) were treated with trichostatin A (5 micromolar for 4 hrs) or 10 micromolar sodium butyrate for 3 hrs before harvesting and preparing for lysates by treating with SDS sample buffer (direct lysates). The lysates can be sonicated and heated (boiling) for 5 mins. SDS-PAGE and Western blot may then proceed." This preparation will help disrupt multimers, which may be causing the additional bands of higher MW so please recommend this preparation to the customer. Performing a secondary only control (no primary) will help determine if the non-specific banding is caused by an interaction with the secondary. Also, it may be beneficial to decrease the amount of secondary used. Please suggest to decrease the primary incubation time from O/N to ~1 hr at room temp. Washing of unbound antibodies may be insufficient. Increase wash periods and add 0.5% TWEEN 20 to wash solution. Also increase the concentration of salt. Increase the number of washes. Washes should be performed for at least 30 minutes changing every 5 minutes, then increase salt (500 mMol) and TWEEN 20 (0.5 to 1.0%) Here are other protocol tips to help: -It may help to try loading less protein. We generally recommend loading 10-30ug of protein depending on the samples. -Increase the detergent (i.e. Tween-20) concentration to 0.5% in your blocking buffer. -Finally, bands larger than the target protein may be due to phosphorylation, glycosylation, splicing variants and additional cross-reactivity with known isoforms. I hope that this information is helpful to your customer. Please let me know if they continue to experience difficulties.

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      Question

      I have a question about one of your products: the cabpa antibody (ab15048) According to the data sheet the the immunogen is aa 981-1348. That is not possible. The mouse protein is only around 360aa I would like to know what epitope was used. Thanks in advance

      Read More
      Answer

      Thank you for your enquiry and your patience in awaiting a response. Ab15048 is derived from a recombinant protein correlating to the nucleotide sequence 466-813 of rat CEBPA (GenBank number NM_012524). The on line data sheet has now been updated to reflect this additional information. If you have any further questions then please do not hesitate to get back in touch with us.

      Read More

      Question

      What is the immunogen? 981-1348 is not correct. The protein is smaller than this!

      Read More
      Answer

      Regarding ab15048, the immunogen used to generate this antibody was rat CEBP alpha recombinant protein, derived from the nucleotide sequence 466-813 of rat origin (GenBank number NM_012524). If you have any additional questions, please contact us again.

      Read More

      Question

      What is the immunogen? 981-1348 is not correct. The protein is smaller than this!

      Read More
      Answer

      Thank you for your enquiry and patience. Yes, it appears that aa 981-1348 is wrong as the CEBP alpha protein is smaller than this. However, I am still waiting to obtain information regarding this from the originator of this antibody. I will contact you again once I receive more information. Thank you again for your patience.

      Read More

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