![]() Always prepare fresh stripping buffer working solution for each blot that will be stripped.ĭiscard the stripping buffer working solution once further stripping of the membrane is no longer needed. ![]() If results are inadequate after completing the protocol, stripping buffer incubation can be carried out at 37 ☌ using a water bath or warm air incubator. Increasing the temperature can significantly improve stripping effectiveness but can also have a highly detrimental effect on reprobing.The Optimization Protocol can help determine the appropriate stripping concentration for the blot and improve the removal of fluorescent signal. Increasing the stripping time and the stripping buffer concentration will progressively reduce the ability to successfully reprobe the blot due to the increased likelihood of damage to the target antigen.Typically, a membrane can be stripped up to three times. The number of times a membrane can be stripped is also dependent on other factors, such as the type and amount of bound antigen, the type of membrane used, and the stringency of stripping conditions.Stripping efficacy on Odyssey PVDF membranes and the number of times a membrane can be stripped are primarily affected by time, concentration, and temperature.Ensure that you have the necessary supplies and reagents for stripping the membrane. NewBlot PVDF 5X Stripping Buffer has not been tested for use with chemiluminescent detection or colorimetric substrates.įamiliarize yourself with the entire document before proceeding.LI-COR neither guarantees nor supports the use of NewBlot PVDF 5X Stripping Buffer with PVDF membranes or antibody-dye conjugates from other manufacturers.NewBlot PVDF 5X Stripping Buffer has been optimized for use with IRDye® 800CW, IRDye 680LT, and IRDye 680RD conjugates.NewBlot PVDF 5X Stripping Buffer has been optimized for use with IRDye antibody conjugates and Millipore Immobilon-FL PVDF membrane.Completed Western blot to be stripped (wet).Large incubation box, such as LI-COR® Incubation Box (11.75 x 8.9 cm, P/N 929-97301).Size: 100 mL (for up to 3,000 cm 2 or approximately fifty 7 x 8.5 cm Millipore Immobilon®-FL PVDF membranes).NewBlot PVDF 5X Stripping Buffer (P/N 928-40032): 100 mL Specifications NewBlot™ PVDF 5X Stripping Buffer Components
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |