Discover Our Latest Application Notes & Protocols



Antibody Reuse: Antibody Recovery and Reuse With the SNAP i.d.® Protein Detection System

Recovering the primary antibody for possible reuse can be very valuable for Western blotting experiments. Here, we show that greater than 90% of the primary antibody can be recovered after the incubation step by following the recommended protocol. The antibodies collected in the SNAP i.d.® 2.0 System can be used successfully in subsequent immunodetection with no reduction in blot quality, even after extended incubations.
Western Blotting Application Notes - Comparison of blots probed with fresh and re-used antibodies in double well blot holders using the SNAP i.d. system
Comparison of cell lysate blots probed with fresh and reused antibodies using the SNAP i.d.® 2.0 system. The blots were probed with anti-PP2A, which was either freshly diluted or recovered and reused, following the SNAP i.d.® 2.0 system extended protocol. Click image to enlarge.


Triple Well Blotting: Triple Well Blot Holder as a Tool for 30-Minute Western Blotting Optimization with the SNAP i.d.® Protein Detection System

Triple Well Blotting: Optimization of antibody concentration is a key requirement for successful immunodetection. Because of the wide range of antibody avidities/affinities, all antibody manufacturers recommend optimization whenever a new antibody is first used in an application. The goal of this effort is to ensure that immunodetection is linear over a defined experimental range as well as to enable the routine production of publication quality Western blots with little or no non-specific background signals.
Western Blotting Application Notes - Replicate blots of whole-cell lysates (lanes 1 – 3: 20, 10 and 5 μg/lane) prepared from human epidermoid carcinoma (A431) cells were probed with a monoclonal antibody specific for the epidermal growth factor receptor diluted from 1:100 to 1:500 as shown
Replicate blots of whole-cell lysates (lanes 1 – 3: 20, 10 and 5 μg/lane) prepared from human epidermoid carcinoma (A431) cells were probed with a monoclonal antibody specific for the epidermal growth factor receptor diluted from 1:100 to 1:500 as shown. The HRP-conjugated goat anti-mouse secondary antibody was used at a 1:10,000 dilution. Click image to enlarge.


Immunodetection: Key Steps for Successful Immunodetection Using the SNAP i.d.® Protein Detection System

Triple Well Blotting: Optimization of antibody concentration is a key requirement for successful immunodetection. Because of the wide range of antibody avidities/affinities, all antibody manufacturers recommend optimization whenever a new antibody is first used in an application. The goal of this effort is to ensure that immunodetection is linear over a defined experimental range as well as to enable the routine production of publication quality western blots with little or no non-specific background signals.

The Western blot is a powerful tool used extensively in protein research to detect and compare the relative levels of proteins without the need for their prior purification. Its widespread appeal is based on its overall simplicity, coupled with the high resolution of proteins separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) prior to membrane transfer.
Western Blotting Application Notes - The SNAP i.d. Protein Detection System has been developed to shorten the time required for immunodetection (i.e., blocking, washing and antibody incubations) to approximately 30 minutes

The SNAP i.d.® Protein Detection System has been developed to shorten the time required for immunodetection (i.e., blocking, washing and antibody incubations) to approximately 30 minutes. Click image to enlarge.


Fluorescent Immunodetection: Fluorescent Immunodetection Using the SNAP i.d.® Protein Detection System

Recent advances in fluorescent dye chemistry and blotting membranes coupled with improvements in instrumentation have accelerated the application of fluorescence detection methodologies to Western blotting. The SNAP i.d.® Protein Detection System has been developed to substantially shorten the time required for fluorescent immunodetection through the use of vacuum.
Western Blotting Application Notes - Cy3 fluorescent immunodetection of GAP43 in human brain lysate. The blot was visualized using Fujifilm FLA_5100 imaging system and was quantified with an R2 of 0.98
Cy3 fluorescent immunodetection of GAP43 in human brain lysate. The blot was visualized using Fujifilm FLA_5100 imaging system and was quantified with an R2 of 0.98. Click image to enlarge.