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Abcam’s Immunoassay Blocking (BSA Free) (ab171535) maintains the activity of antibodies and other biomolecules adsorbed or immobilized to solid substrates. It is easily incorporated into most assay protocols as the blocking solution, and minimal incubation is required. For blocking nonspecific sites on hydrophobic substrates, incubate for as little as five minutes and then proceed with your assay. Longer incubation times, while not necessary, are also not detrimental. For simultaneous blocking and stabilizing, apply ab171535 and incubate. Remove excess solution and completely dry the component. For maximum shelf life, store the stabilized component in a sealed container with desiccant.
To Stabilize Adsorbed or Immobilized Proteins on Microtiter Plates/Strips
1. For optimal stability and performance, use ab171535 at 100% concentration
2. 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 material
3. Immediately after washing, add ab171535 to allow interaction with the entire protein-coated surface; do not let coated components dry before adding ab171535 since drying contributes to the loss of protein activity
4. Incubate for five minutes to two hours at room temperature; for most assays, ab171535 can replace the blocking solution
5. Remove or aspirate ab171535, but do not wash
6. To dry components for long-term storage, we recommend one of the following methods:
– Option 1. Place plates in a humidity-controlled chamber until dry (2-24 hours)
– Option 2. Dry 2 -24 hours in a sealed desiccated container, such as a cabinet, jar, or plastic bag containing calcium sulfate or silica gel desiccant in amounts adequate for the size of the container, or
– Option 3. Dry plates at 30°C-40°C in a vacuum oven for 2-4 hours
NOTE: Drying times should be optimized for each application
7. Immediately package the dried product in an airtight, moisture-proof container with a desiccant; this is especially important with the final product is stored in a humid environment or refrigerated where condensation is likely to occur
Our Abpromise guarantee covers the use of ab171535 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent concentration.|
Stability tests were performed with Immunoassay Blocking (BSA Free) (ab171535) using either monoclonal or polyclonal IgG antibodies.
Coated microtiter plates were stabilized with either ab171535 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 (BSA Free) (ab171535) at room temperature, the monoclonal antibody retained greater than 85% activity and the polyclonal antibody retained 90% activity.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"