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Biotinylated Goat Anti-Rabbit IgG (H+L) (Ready to use) (ab64256)

Reviews (1)Q&A (3)References (103)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotinylated Goat Anti-Rabbit IgG (H+L) (Ready to use) (ab64256)

    製品の概要

    • 製品名

      Biotinylated Goat Anti-Rabbit IgG (H+L) (Ready to use)
      IgG ihc 試薬 製品一覧

    • アプリケーション

      適用あり: IHC-Pmore details

    • 特記事項

      Staining protocol
      1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.
      2. Add appropriate hydrogen peroxide blocking solution to cover the sections. Incubate for 10 minutes. Wash 2 times in buffer.
      3. Perform appropriate pretreatment if required. Wash 3 times in buffer.
      4. Apply appropriate protein blocking solution and incubate for 5 minutes at room temperature to block non specific background staining. Wash 1 time in buffer.
      5. Apply primary antibody and incubate according to manufacturer's protocol.
      6. Wash 4 times in buffer. Apply Ready to use Biotinylated Goat anti Rabbit IgG (ab64256) solution and incubate for 10-15 minutes at room temperature. Wash 4 times in buffer.
      7. Apply enzyme labelled streptavidin and incubate according to manufacturer’s protocol.
      8. Rinse 4 times in buffer. Place slide in appropriate chromogenic substrate and incubate until desired reaction is achieved. Rinse 4 times in buffer.
      9. Add enough drops of Haematoxylin to cover the section. Incubate for 1 minute.
      10. Rinse 7-8 times in tap water. Add Mounting Medium to cover the section.


      This reagent is intended to be used in a labeled streptavidin biotin immunoenzymatic antigen detection system. This technique involves the sequential incubation of the specimen with an unconjugated primary antibody specific to the target antigen, a biotinylated secondary antibody which reacts with the primary antibody, enzyme labeled streptavidin, and substrate chromogen.

    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ab64256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 at an assay dependent dilution.
    特記事項
    IHC-P
    Use at an assay dependent dilution.

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotinylated Goat Anti-Rabbit IgG (H+L) (Ready to use) (ab64256)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotinylated Goat Anti-Rabbit IgG (H+L) (Ready to use) (ab64256)
      Ab64256 was used with the primary antibody ab3517 in IHC-P. See Abreview on ab3517.

    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 (103)

    ab64256 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。

    ab64256 は 103 報の論文で使用されています。

    • Rusin D  et al. Microglia-Derived Insulin-like Growth Factor 1 Is Critical for Neurodevelopment. Cells 13:N/A (2024). PubMed: 38247874
    • Pakkarato S  et al. Discrete localization patterns of PIP5Kγ and PLCβ3 working sequentially in phosphoinositide-cycle within mouse sensory neuron somata. Microsc Res Tech 86:351-358 (2023). PubMed: 36579633
    • Rämö JT  et al. Genome-wide screen of otosclerosis in population biobanks: 27 loci and shared associations with skeletal structure. Nat Commun 14:157 (2023). PubMed: 36653343
    • Ashraf UM  et al. Intrarenal Dopaminergic System Is Dysregulated in SS-Resp18mutant Rats. Biomedicines 11:N/A (2023). PubMed: 36672619
    • Zhang Y  et al. Copper Induces Cognitive Impairment in Mice via Modulation of Cuproptosis and CREB Signaling. Nutrients 15:N/A (2023). PubMed: 36839332
    View all Publications for this product

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    1-4 of 4 Abreviews or Q&A

    Specific, Easy IHC secondary anti rabbit antibody

     Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    abreview image
    I have been using Abcam products for most of my experiments with western blotting and IHC. In particular, this secondary biotinylated antibody was used for IHC. This antibody is easy to use and more specific towards primary antibody. I have used in particular to stain BCRP-1 protein in the human placenta. This secondary is specific and stains intercellularly.
    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 May 03 2021

    Question

    How is this antibody biotinylated? Is it biotinylated at just the N terminus, at all amines, or other? Is the ratio of bitoin to antibody known? If not can you at least provide an estimate? What is the concentration of the antibody?

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    Answer

    Thank you for contacting us about the Biotinylated Goat Anti Rabbit IgG antibody, ab64256.

    The ratio of biotin to antibody is estimated to be 6 - 8. The concentration of the antibody is 5ug/ml.

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

    Read More

    Question

    I need to develop double IHC analysis on formalin-fixed paraffin-embedded (FFPE) tissue sections of human tumors specimens, and after we have to do laser capture microdissection of positive cells.   I already have two primary antibodies directed against CD44 (produced in mouse) and against ALDH1 (produced in rabbit).   I'm looking for double staining kit (or all of the individual components which analysis needs) with alcohol insoluble chromogens because after IHC and before laser cature microdissection I have to dehydrate the slides.   Thank you for your time and consideration.

    Read More
    Answer

    Thank you for contacting us.

    We do not currently offer double staining IHC kits. Your staining conditions are challenging as, of the two common HRP chromogens, only DAB is alcohol insoluable. While of the common AP chromagens, only BCIP/NBT is insoluable. This means that to do double labeling you will have to do a sequential stain first using one detection system and then with the other.

    I have collected a list, with links, of the reagents which you would need for this type of staining.

    1) After de-parrafinizing and re-hydrating the samples, perform an antigen retrieval step treating with Cirate buffer pH6.0,https://www.abcam.com/10x-Citrate-Buffer-pH-6-0-ab64214.html, in the microwave or pressure cooker. After performing the antigen retrieval rinse in cold tap water for 10 minutes.

    2) Rinse slides in TBST,https://www.abcam.com/20x-TBS-T-with-Tween-20-ab64250.html.

    3) Performing a blocking step using protein block,https://www.abcam.com/Protein-Block-ab64226.html.

    4) Incubate sections with your first primary antibody, for clarity I will assume the CD44 antibody, raised in mouse.

    5) Wash with TBST.

    6) Block endogenous peroxidases using aHydrogen Peroxide Blocking Reagent,https://www.abcam.com/Hydrogen-Peroxide-Blocking-Reagent-ab94666.html.

    7) Incubate your tissue with a biotinylated secondary antibody, in this instance anti-Mouse IgG,https://www.abcam.com/Biotinylated-Goat-anti-Mouse-IgG-H-L-Ready-to-Use-ab64255.html.

    8) Wash with TBST.

    9) Detect your first antibody usingStreptavidin Peroxidase (Ready to Use)https://www.abcam.com/Streptavidin-Peroxidase-Ready-to-Use-ab64269.html. Note that this does not containa serum or BSA solution as diluent. Serum may contain biotin therefore competing Streptavidin binding with biotinylated secondary antibody, therefore reducing binding activity.

    10) Apply your chromogen, DABsubtrate,https://www.abcam.com/DAB-Substrate-Kit-ab64238.html.

    11) Wash with TBST.

    12)Performing a second blocking step using protein block,https://www.abcam.com/Protein-Block-ab64226.html.

    13)Wash with TBST.

    14)Incubate sections with the your second primary antibody, this time ALDH1, raised in rabbit.

    15)Wash with TBST.

    16)Incubate your tissue with a biotinylated secondary antibody, in this instance anti-Rabbit IgG,https://www.abcam.com/Biotinylated-Goat-Anti-Rabbit-IgG-H-L-Ready-to-use-ab64256.html

    17)Wash with TBST.

    18)Detect your first antibody usingStreptavidin Alkaline Phosphatase (Ready to Use)https://www.abcam.com/Streptavidin-Alkaline-Phosphatase-Ready-to-Use-ab64268.html.

    19) Apply your second chromogenAlkaline Phosphatase chromogen (BCIP/NBT) - Ready to Use,https://www.abcam.com/Alkaline-Phosphatase-chromogen-BCIP-NBT-Ready-to-Use-ab7468.html.

    18) Wash with TBST, counterstain if desired.


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

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    Question

    Hello, I just wanted to confirm that this antibody has the ability to recognize both "mouse and rabbit" regions and can be used as secondary antibody for recognizing Mouse Antibodies and Rabbit Antibodies. Also is there any technical specifications as compare to individual secondary antibodies ab64256 (Anti-rabbit) and ab97044 (Anti-mouse)? Let me know as soon as possible. Thanks and Warm Regards,

    Read More
    Answer

    Thank you for your inquiry. ab64257 is indeed suitable for the detection of Mouse and Rabbit IgG. It can be used as a secondary antibody when used with rabbit or mouse primary antibodies. About ab64256 and ab97044, all of the information we have for each of our products is listed on the online datasheet for your convenience. These are updated as soon as any new information is brought to our attention. Please do not hesitate to contact us again if you need further assistance.

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