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 hours6.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
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.
|ELISA||Use at an assay dependent concentration.|
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.