#Ez ladder protein western blot serial#

#Ez ladder protein western blot serial#

Perform serial dilutions of the lysate or sample to determine the optimal amount of protein to load.Too little lysate leads to insufficient availability of the target of interest.Insufficient quantity of sample loaded on the gel Dilute your primary antibody in wash buffer.Some blocking solutions can mask the blot and reduce the availability of the antigen to the antibody, especially if the blocking step is >1 hour.Too much detergent or the nature of the detergent can result in washing away the signal-decrease or eliminate detergent.Ensure the correct excitation and emission ranges are selected for the intended fluorophore.Use the Smart Exposure feature on the iBright FL1500 system, or a comparable autoexposure feature on another instrument.Avoid multiple uses of pre-diluted antibodies.Check the expiration date of the antibody.

Ensure the antibody was stored appropriately.Extend the incubation time to overnight at 4☌, or 3–6 hours at room temperature.Ensure primary antibody has a good titer and is specific for the antigen to be detected.Increase primary antibody concentration.Spectra Multicolor Low Range Protein Ladder Spectra Multicolor High Range Protein Ladder NuPAGE MOPS SDS Buffer (15–260 kDa) Bolt MES SDS Buffer (3.5–160 kDa) add Bolt Antioxidant for reduced samples Low-abundance/ posttranslationally modifiedīolt MOPS SDS Buffer (15–260 kDa) Bolt MES SDS Buffer (3.5–160 kDa) add Bolt Antioxidant for reduced samples