Anti-Bim 抗体 (ab7888)

Submit a review Q&A (5)References (19)
Western blot - Anti-Bim antibody (ab7888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bim antibody (ab7888)
  • Western blot - Anti-Bim antibody (ab7888)
  • Immunocytochemistry/ Immunofluorescence - Anti-Bim antibody (ab7888)
  • Immunocytochemistry/ Immunofluorescence - Anti-Bim antibody (ab7888)

Key features and details

  • Rabbit polyclonal to Bim
  • Suitable for: IHC-P, WB, ICC/IF
  • Reacts with: Rat, 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-Bim antibody [Y36] (ab32158)
  • 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-Bim antibody
    Bim 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to Bim

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IHC-P, WB, ICC/IFmore details

  • 種交差性

    交差種: Rat, Human

  • 免疫原

    Peptide corresponding to amino acids 22 to 40 of human origin. The sequence is identical to that of mouse and differs from that of rat by one amino acid.

  • ポジティブ･コントロール

    • WB: Human colon and thymus tissue lysates, Rat Myeloma Cell lysate ICC/IF: K562 cells

  • 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ab7888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
Use a concentration of 20 µg/ml.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 23 kDa.
ICC/IF
Use at an assay dependent concentration.
特記事項
IHC-P
Use a concentration of 20 µg/ml.
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 23 kDa.
ICC/IF
Use at an assay dependent concentration.

ターゲット情報

  • 機能

    Induces apoptosis. Isoform BimL is more potent than isoform BimEL. Isoform Bim-alpha1, isoform Bim-alpha2 and isoform Bim-alpha3 induce apoptosis, although less potent than the isoforms BimEL, BimL and BimS. Isoform Bim-gamma induces apoptosis.

  • 組織特異性

    Isoform BimEL, isoform BimL and isoform BimS are the predominant isoforms and are ubiquitously expressed with a tissue-specific variation. Isoform Bim-gamma is most abundantly expressed in small intestine and colon, and in lower levels in spleen, prostate, testis, heart, liver and kidney.

  • 配列類似性

    Belongs to the Bcl-2 family.

  • ドメイン

    The BH3 motif is required for Bcl-2 binding and cytotoxicity.

  • 細胞内局在

    Mitochondrion and Endomembrane system. Associated with intracytoplasmic membranes.
  • Information by UniProt

  • 参照データベース

    • 別名

      • BCL2 like 11 antibody
      • B2L11_HUMAN antibody
      • BAM antibody
      • Bcl 2 interacting protein Bim antibody
      • Bcl 2 related ovarian death agonist antibody
      • Bcl-2-like protein 11 antibody
      • BCL2 interacting mediator of cell death antibody
      • BCL2 like 11 (apoptosis facilitator) antibody
      • BCL2 like protein 11 antibody
      • Bcl2-interacting mediator of cell death antibody
      • Bcl2-L-11 antibody
      • Bcl2l11 antibody
      • BIM alpha6 antibody
      • BIM antibody
      • BIM beta6 antibody
      • BIM beta7 antibody
      • BimEL antibody
      • BimL antibody
      • BOD antibody
      see all

    画像

    • Western blot - Anti-Bim antibody (ab7888)
      Western blot - Anti-Bim antibody (ab7888)
      All lanes : Anti-Bim antibody (ab7888) at 0.5 µg/ml

      Lane 1 : Human thymus tissue lysate
      Lane 2 : Human colon tissue lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

      Predicted band size: 23 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bim antibody (ab7888)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bim antibody (ab7888)

      ab7888 at 20µg/ml staining Bim in human cancer cells by IHC-P

    • Western blot - Anti-Bim antibody (ab7888)
      Western blot - Anti-Bim antibody (ab7888)
      Anti-Bim antibody (ab7888) at 0.5 µg/ml + Rat Myeloma Cell lysate at 15 µg

      Secondary
      Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

      Predicted band size: 23 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-Bim antibody (ab7888)
      Immunocytochemistry/ Immunofluorescence - Anti-Bim antibody (ab7888)
      Immunocytochemistry/ Immunofluorescence analysis of 4% paraformaldehyde fixed human K562 cells labeling Bim with ab7888 at 20 μg/mL. Goat anti-rabbit IgG secondary antibody at 1/500 dilution was used as the secondry antibody.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-Bim antibody (ab7888)
      Immunocytochemistry/ Immunofluorescence - Anti-Bim antibody (ab7888)
      ICC/IF image of ab7888 stained HepG2 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7888, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    参考文献 (19)

    ab7888 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。

    ab7888 は 19 報の論文で使用されています。

    • Song J  et al. The 14-3-3s protein promotes HCC anoikis resistance by inhibiting EGFR degradation and thereby activating the EGFR-dependent ERK1/2 signaling pathway. Theranostics 11:996-1015 (2021). PubMed: 33391517
    • Liu XK  et al. Regulatory Effect of Sishen Pill on Tfh Cells in Mice With Experimental Colitis. Front Physiol 11:589 (2020). PubMed: 32581849
    • Zhang J  et al. MicroRNA-132 protects H9c2 cells against oxygen and glucose deprivation-evoked injury by targeting FOXO3A. J Cell Physiol 235:176-184 (2020). PubMed: 31210352
    • Mei X  et al. lncRNA-NR024118 overexpression reverses LPS-induced inflammatory injury and apoptosis via NF-?B/Nrf2 signaling in ATDC5 chondrocytes. Mol Med Rep 20:3867-3873 (2019). PubMed: 31485657
    • Wang Y  et al. Necrostatin-1 ameliorates the pathogenesis of experimental autoimmune encephalomyelitis by suppressing apoptosis and necroptosis of oligodendrocyte precursor cells. Exp Ther Med 18:4113-4119 (2019). PubMed: 31611942
    View all Publications for this product

    レビューと Q&A

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    1-5 of 5 Abreviews or Q&A

    Question

    Thanks for your reply. I had beta tubulin as the loading control which worked perfectly fine. So I do not think there is a problem with the cell lysate. I stored the antibody at -20 degree celsius afeter alliquoting it in order to avoid repeated freeze/thaw cycles. Unfortunately I do not have access to K562 or A549 whole cell lysates. I did an overnight incubation with the primary antibody at 4 degree celsius. I saw on your website that you also have a monoclonal antibody against BIM ( ab32158). I was wondering if you could send me that antibody ie Anti-BIm antibody (Y36) catalog number ab32158 as a replacement so that I can test it on my samples.

    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 of ab32158.

    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

    Amount of protein loaded was 100ug. Blocked with 5% milk. Primary used 1:1000 in 5% milk. Secondary used 1:2000 in 5% milk. Transfer done using i blot. Washing done 3 times with each wash 15 minutes. Previously I had ordered this antibody in September and it worked a couple of times and then stopped working. I ordered this again last week and keeping exactly the same conditions as before tried it again but it did not work. Samples used were HEK 293, TC32 and SKES cell lysates. The two lot numbers are GR65948-2 and GR85357-1 . I am not sure which lot came last week and which came in September. The one which came in September worked a couple of times while the one which came last week did not work at all. Can you check which of the above lots you sent me in September and send it to me again.

    Read More
    Answer

    Thank you for taking time to contact us. I am sorry to hear that this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Would you be able to clarify a few points for me?

    When you say that the antibody wasn't working, was the issue that you were not seeing any bands at all? Have you probed the same membranes for antibodies to other targets (i.e. loading control)?

    Do you have access to K562 or A549 whole cell lysates to use as a positive control?

    Was the primary antibody incubated overnight at 4C?

    How was the antibody stored?

    I took a look at our stocks and unfortunately we do not have any more of the lot you purchased in September (GR85357-1). We only have lot GR65948-2 in stock now.

    In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. Please let me know how you would like to proceed.

    I hope this information is helpful, and I thank you for your cooperation.

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    Question

    The samples were prepared using the attached protocol. 30ug of protein was loaded on the gel. I have attached an improved image of the western blot image with labelled molecular weight markers.

    I blocked the membrane that was incubated with Gadd153 for 1h only.

    Also, the secondary antibodies that I made were fresh but the aliquot I used, was stored at 4 degrees C. Maybe I should try using a fresh aliquot from -20 degrees Celsius. It could be due to the secondary antibody. I will be repeating this procedure tomorrow or sometime this week just to confirm the results.

    I would really appreciate it if you could assist me with any changes that I need to make to my existing protocol to get better results.

    Thank you very much.

    Read More
    Answer

    Thank you for your reply.


    I am inclined to believe that the issue is with the samples themselves. It is very unusual for 3 different antibodies to detect bands mainly above 100kDa when the expected MW should be around 25kDa. How long are the samples boiled before loading? In reading your protocol, I could only find 1 minute, which is not sufficient. Boiling 5-10 min is advised to completely denature the proteins.


    How do the samples look if you stain the gel with Coomassie? or Ponceau S?

    Furthermore, for at least GADD153, expression must be induced in your samples. Did you do this? Also, ab54740 is only validated for human and may not work with your MEFs. Are you running any sort of a loading control to confirm that the procedure is working with other antibodies?

    I hope this information is helpful. Please do not hesitate to contact me if you have additional questions.

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    Question

    I have attached the image of the blot and I have tried to provide you with the description of my experiment. I used Mouse Embryonic Fibroblast (MEF) cells H2AX+/+ and H2AX-/- for my study. I used the same samples to test all three antibodies and cut it to incubate them separately in different antibodies. I have also attached the protocol that I used to perform western blot.

    Thank you very much for your assistance.

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    Answer

    Thank you for your reply.

    I was wondering if you would clarify a few additional points for me.


    How were the samples prepared? How much is loaded on the gel?


    What is the molecular weight of each band in the ladder you provided?


    Regarding the WB protocol, each protein may have slightly different requirements, so it is important to consider these when interpreting your results. For example, GADD153 must be induced to be detected by WB with this antibody - how did you treat your cells? Additionally, background is expected with this antibody and a 2hr block with milk is recommended.


    Can you confirm if the secondary antibody you have used with these antibodies is made fresh? Your protocol indicated that your lab does reuse secondary antibody, but we do not recommend it. Have you performed no primary controls to be sure your results are due to the primary antibodies?


    I look forward to your reply so that I may assist you further. I think some modifications to your protocol will help your results with these antibodies. Please do not hesitate to contact me if you have any additional questions.

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    Question

    We ordered 3 primary antibodies- Bim, Gadd45a and Gadd153. I used these to test them and my blot shows unspecific binding of antibodies. I was wondering if there is any information that you can provide me with to interpret these results or if there is anyone I could contact?
    Thank you very much.

    Read More
    Answer

    Thank you for contacting Abcam.


    I am sorry that you have been experiencing difficulties with these antibodies in WB.


    In order to assist you further, I am hoping you can provide some additional details regarding your experiments. Would you please send me a labeled image of your blot, detailed protocol, and sample type and preparation information? Once I have these details, I would be happy to offer some protocol suggestions to improve your results or offer compensation in accordance with our Abpromise.


    I look forward to your reply. Please do not hesitate to contact us if you have any additional questions.

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