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    products/reagents/10x-blocking-buffer-ab126587.pdf

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10X Blocking Buffer (ab126587)

  • Datasheet
  • SDS
Submit a review Q&A (4)References (18)

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10X Blocking Buffer (ab126587)

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    Overview

    • Product name

      10X Blocking Buffer
    • Tested applications

      Suitable for: WB, ELISA, In-Cell ELISA, ICC/IFmore details
      Unsuitable for: Flow Cyt
    • General notes

      Protein blocking buffer (pH 7.5) (ab126587) for serum-free blocking of non-specific antibody binding in ICC, ELISA and western blot. BSA-free

      Preparation: Dilute to 1X in deionized water or PBS (e.g. 1mL 10X blocking buffer + 9mL water/PBS). Diluting in PBS instead of water makes a somewhat more stringent blocking buffer due to the presence of salt in the PBS. Blocking Buffer is pH 7.5. The user should determine empirically whether dilution in water or PBS is most appropriate for the assay at hand. Use diluted block solution promptly. 

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Store at Room Temperature. The product can be stored for up to 12 months.
    • Storage buffer

      Constituent: Proprietary component
    • Concentration information loading...
    • Research areas

      • Kits/ Lysates/ Other
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      • IHC Tools/ Reagents
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      • Serum Products
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      • Western Blot Tools and Reagents
      • Optiblot buffers
      • Kits/ Lysates/ Other
      • Kits
      • IHC tools
      • Block
      • Protein Block

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab126587 in the following tested applications.

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

    Application Abreviews Notes
    WB
    Use at an assay dependent concentration.
    ELISA
    Use at an assay dependent concentration.
    In-Cell ELISA
    Use at an assay dependent concentration.
    ICC/IF
    Use at an assay dependent concentration.
    Notes
    WB
    Use at an assay dependent concentration.
    ELISA
    Use at an assay dependent concentration.
    In-Cell ELISA
    Use at an assay dependent concentration.
    ICC/IF
    Use at an assay dependent concentration.
    Application notes
    Is unsuitable for Flow Cyt.

    Images

    • 10X Blocking Buffer (ab126587)
      10X Blocking Buffer (ab126587)

      Primary :

      All Lanes : top membranes with Anti AMPK pThr172 + Thr183 (ab133448) at 1/1000. Bottom membranes with Anti AMPKa (ab110036) at 1/1000

      Lane 1 = AMPKa1 protein

      Lane 2 = C2C12 cells treated with AICAR

      Lane 3 = untreated C2C12 cells

      Secondary: Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) and Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1:10,000 developed using the ECL technique.

      Performed under reducing conditions (50mM DTT – Sample heated at 60ºC). Predicted band size for AMPKa: 64 kDa Observed band size for AMPKa: 64 kDa

      Left membrane All blocking steps = 1X blocking reagent ab126587 diluted in PBST

      Right membrane All blocking steps = 5% Milk in PBST Exposure time : 5 minutes



      Primary :

      All Lanes : top membranes with Anti AMPK pThr172 + Thr183 (ab133448) at 1/1000. Bottom membranes with Anti AMPKa (ab110036) at 1/1000

      Lane 1 = AMPKa1 protein

      Lane 2 = C2C12 cells treated with AICAR

      Lane 3 = untreated C2C12 cells

      Secondary: Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) and Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1:10,000 developed using the ECL technique.

      Performed under reducing conditions (50mM DTT – Sample heated at 60ºC). Predicted band size for AMPKa: 64 kDa Observed band size for AMPKa: 64 kDa

      Left membrane All blocking steps = 1X blocking reagent ab126587 diluted in PBST

      Right membrane All blocking steps = 5% Milk in PBST Exposure time : 5 minutes

    Protocols

    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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (18)

    Publishing research using ab126587? Please let us know so that we can cite the reference in this datasheet.

    ab126587 has been referenced in 18 publications.

    • Yang JM  et al. Gradient Waveguide Thickness Guided-Mode Resonance Biosensor. Sensors (Basel) 21:N/A (2021). PubMed: 33430392
    • Dang TM  et al. Optimization of the isolation procedure and culturing conditions for hepatic stellate cells obtained from mouse. Biosci Rep 41:N/A (2021). PubMed: 33350435
    • Yokoe I  et al. Lipid bubbles combined with low-intensity ultrasound enhance the intratumoral accumulation and antitumor effect of pegylated liposomal doxorubicin in vivo. Drug Deliv 28:530-541 (2021). PubMed: 33685314
    • Ho CNQ  et al. Simulated Microgravity Inhibits the Proliferation of Chang Liver Cells by Attenuation of the Major Cell Cycle Regulators and Cytoskeletal Proteins. Int J Mol Sci 22:N/A (2021). PubMed: 33925309
    • Chou LY  et al. Discoidin Domain Receptor 1 Regulates Runx2 during Osteogenesis of Osteoblasts and Promotes Bone Ossification via Phosphorylation of p38. Int J Mol Sci 21:N/A (2020). PubMed: 33003599
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Question

    Is there BSA in this product?

    Read More

    Abcam community

    Verified customer

    Asked on Jul 16 2013

    Answer

    The the lab has confirmed this blocking solution is casein based with no BSA, so it may be an option for your ELISA if this is a concern for you.

    Please let me know if you have any further questions!

    Read More

    Heather Allen

    Abcam Scientific Support

    Answered on Jul 17 2013

    Question

    can this buffer also be diluted with TBS?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 19 2013

    Answer

    Thank you for contacting us.



    Yes – in our experience, as with PBS, the extra salt in TBS will make a more stringent block/ prevent non specific binding.





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

    Read More

    Abcam Scientific Support

    Answered on Mar 19 2013

    Question

    Customer kindly contacted us regarding our 10X blocking buffer ab126587. Specifically whether this is the buffer present in kit ab111542 and the %casein in the blocking solution. Additionally, the customer was wondering what the composition of the blocking buffer in ab126420 is.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 21 2012

    Answer


    1. ab126420, the components of the blocking buffer are proprietary, and the solution is a standard blocking solution.

    2. ab111542: The blocking solution in the kit is the same as the 10X blocking solution sold separately (ab126587). It is casein based and the % of casein is proprietary.

    Read More

    Abcam Scientific Support

    Answered on Dec 21 2012

    Question

    I ordered ab135575 and want to do ICC/IF as shown on website and antibody literature (with HEK293). What did you use to block? Since this is an albumin antibody, I don't think 1%BSA or serum is appropriate. Thank you.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 14 2012

    Answer

    Thank you for contacting us.

    In this case a casein block was used as BSA and serums would cause high background. We do carry a casein based blocking solution, ab126587,which we have used extensively and feel is a superior product in ICC. More information about ab126587 may be found at the following link:

    https://www.abcam.com/10x-blocking-buffer-ab126587.html

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

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Aug 14 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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