Overview

  • Product name
    Immunoassay Blocking Buffer
    See all ELISA buffer reagents
  • Tested applications
    Suitable for: ELISAmore details
    Unsuitable for: IHC-Fr or IHC-P
  • General notes

    Abcam’s Immunoassay Blocking Buffer (ab171534) effectively preserves the conformation and activity of dried proteins in immunoassays. Immunoassay Blocking Buffer simultaneously blocks and stabilizes with superior results. It can be used to stabilize antibodies, antigens or enzymes on an assortment of immunoassay components, such as polystyrene plates, tubes, glass, membranes, and filter paper. It is easily incorporated into most assay protocols by simply substituting it for the blocking solution.

    Recommendations for Use: The following are general guidelines only.

    To Stabilize Adsorbed or Immobilized Proteins on Microtiter Plates/Strips
    1.Immobilize or adsorb the primary protein (antibody or antigen) according to the procedure optimized in your laboratory; wash adequately to remove excess or weakly bound protein
    2.Immediately after washing, add Immunoassay Blocking Buffer stabilizer to allow interaction with the entire protein-coated surface; do not let the components dry before adding stabilizer since drying contributes to the loss of protein activity
    3.Incubate 15-60 minutes at room temperatures; for most assays, Immunoassay Blocking Buffer stabilizer can replace the blocking solution
    4.Remove or aspirate the Immunoassay Blocking Buffer stabilizer, but do not wash
    5.Dry components for long-term storage. Wells coated with Immunoassay Blocking Buffer stabilizer may require longer drying times than those without; drying times should be optimized for each application. Recommended methods are:

    - Place plates in a humidity-controlled chamber until dry (4-24 hours)

    - Dry plates at 30°C-40°C in a vacuum oven for 4 hours

    6.Package the final, stabilized plates/strips in an airtight container with desiccant

    To Stabilize Adsorbed or Immobilized Proteins on Membranes
    1. Immobilize or adsorb the primary protein (antibody or antigen) according to the procedure optimized in your laboratory
    2. Dilute 1 part Immunoassay Blocking Buffer stabilizer in three parts compatible buffer or deionized water; add 0.01% surfactant
    3. Coat membrane by incubating or spraying with the Immunoassay Blocking Buffer stabilizer solution
    4. Dry thoroughly. Faster drying results in better flow properties
    5. Package the final, stabilized membrane in an airtight container with a desiccant

    To Stabilize Conjugates in the Dry Form
    1. If you dilute your conjugate before drying/lyophilizing, use Immunoassay Blocking Buffer stabilizer as the diluent. Otherwise, add between 5-10 parts Immunoassay Blocking Buffer stabilizer or 1 and 2 parts 5X Immunoassay Blocking Buffer stabilizer to 1 part conjugate
    2. If you are lyophilizing in vials with rubber stoppers, stability may be improved by placing the rubber stoppers in a 100°C vacuum oven for one hour prior to use; this dries them and drives off any volatiles
    3. Freeze the conjugate/stabilizers mixture, then lyophilize as normal; lyophilization may require extra time
    4. For evaporation drying, place the conjugate/stabilizer mixture in a 37°C-40°C oven for four hours or until completely dry; the volume per container should be low enough to allow maximum surface area to be exposed to air during drying; store the final product in an airtight container

Properties

  • Form
    Liquid
  • Storage instructions
    Store at room temperature.
  • Storage buffer
    Constituent: 100% Proprietary component

    pH 7.0 - 7.4. Contains BSA. Preservative: None.
  • Concentration information loading...

Associated products

Applications

Our Abpromise guarantee covers the use of ab171534 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
ELISA Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-Fr or IHC-P.
  • Images

    • Stability tests were performed with Immunoassay Blocking Buffer (ab171534) using either monoclonal or polyclonal IgG antibodies.

      Coated microtiter plates were stabilized with either ab171534 or 1% BSA. The antibody-coated plates were dried and placed at room temperature. Percent retained activity was determined by comparing the activity of the aged antibody-coated microtiter plates to that of freshly coated plates.

      After 30 months in Immunoassay Blocking Buffer (ab171534) at room temperature, the monoclonal antibody retained 70% activity and the polyclonal antibody retained 95% activity.

    References

    This product has been referenced in:
    • Ahmed SR  et al. Amplified visual immunosensor integrated with nanozyme for ultrasensitive detection of avian influenza virus. Nanotheranostics 1:338-345 (2017). Read more (PubMed: 29071197) »
    See 1 Publication for this product

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

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