Immobilon® GO is a hands-free, immunodetection device designed for mini Western blot processing. Our unique design uses continuous, sequential fluid convection across a flow matrix to enable autonomous antibody binding and membrane wash steps during immunodetection. Immobilon® GO is intended for one-time use and does not require an external power or vacuum source for operation. Standard non-fat dry milk TBST buffer is used to allow for easy transition from traditional Western blot protocols. Western blots can be processed in as little as 3 hours or may remain in the device overnight for processing.
The Immobilon® GO immunodetection device contains two parts: the base assembly with flow matrix (Figure 1) and the device lid (Figure 2).
Figure 1.Components of Immobilon® GO device base assembly
Figure 2.Components of Immobilon® GO device lid
Materials required for immunodetection, but not supplied:
Figure 3.Block membrane by incubating in 2% non-fat dry milk blot buffer
4. Prepare primary and secondary antibody dilution in 2% NFDM blot buffer. A total volume of 1 mL is required for each antibody.
5. Open Immobilon® GO device and fold back both parts of the flow matrix (Figure 4).
Figure 4.Remove lid and open both pieces of the flow matrix
6. Position blotting membrane (protein side up), 1 cm from stage edge. Low molecular weight proteins should be facing the notched flow matrix (Figure 5).
Figure 5.Place blocked membrane 1 cm from the stage edge with low molecular weight proteins towards the loading port
7. Fold flow matrix over the membrane and add 4 mL of 2% NFDM blot buffer evenly across the flow matrix (Figure 6).
Figure 6.Fold top portion of flow matrix over the membrane and add NFDM blocking buffer
8. To assure even contact between the membrane and flow matrix, pull the blot roller from the loading port end toward the flow matrix tab (Figure 7).
Figure 7.Use blot roller to assure even contact between the membrane and flow matrix
9. Using the blot roller fold over the flow matrix tab. Assure all air bubbles have been removed and close contact between the flow matrix and membrane has been established (Figure 8).
Figure 8.Seal flow matrix by folding over flow matrix tab with the blot roller.
10. Replace the lid (Figure 9).
Figure 9.Replace lid on Immobilon® GO device
11. Seal lid by gently push down in area specified by loading ports; a clicking sound indicates that lid is closed (Figure 10).
Figure 10.Seal lid by gently pressing down in designated areas until a clicking sound is heard
12. To activate Immobilon® GO device, add 2% NFDM blot buffer to the reagent loading ports in the order and volumes shown below (i.e., fill port 1A, followed by 1B) (Figure 11).
Figure 11.Addition of buffer and antibodies to loading ports
13. Add the primary and secondary antibodies in the order and volumes as outlined in the table below.
For first time use of the Immobilon® GO immunodetection device, please refer to the detailed user guide.
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