Overview

  • Product name10X Blocking Buffer
  • Tested applicationsSuitable for: ICC, WB, ELISA, In-Cell ELISA, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • General notesDescription: A casein-based blocking solution buffered to pH 7.5. An efficient blocking solution to reduce non-specific background staining in immunoassay techniques including western blot, immunocytochemistry, sandwich ELISA and in-cell ELISA.

    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. The diluted block solution can be stored at 4C for short periods of time.
    Shelf life for the 10X stock is 1 year.

Properties

    Applications

    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
    ICC Use at an assay dependent concentration.
    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 notesIs unsuitable for Flow Cyt.
  • 10X Blocking Buffer images

    • 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

    References for 10X Blocking Buffer (ab126587)

    ab126587 has not yet been referenced specifically in any publications.

    Product Wall

    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!

    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.





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    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. M...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"