Western Blot normalization
Sample quality control
Accurately compare the band strength of different lanes. Precautions
When performing quantitative western blot, it is necessary to compare the intensity values ​​of the bands in different lanes. The premise is to control the sample volume, correct the deviation, and ensure that the sample volume is consistent in each lane.
Typically, researchers calibrate by detecting the amount of housekeeping/structural protein that is consistent in all cases.
For chemiluminescence detection, when both proteins are detected, the blotting membrane needs to be stripped and re-incubated.
One of the great advantages of fluorescent western blot detection is that two proteins can be detected simultaneously, and the sample volume control can be achieved without stripping and secondary incubation.
Fluorescence detection can simultaneously carry out sample quality control, and one blot film detects two colors.
In this experiment, the primary antibody must be derived from two different species, while the secondary antibody corresponds to its species and labeled with different fluorescent labeling groups.
AzureSpectra secondary antibody (sheep) is labeled with an infrared fluorescent group and has a variety of secondary antibodies against rabbits, mice, humans, chickens, rats, and guinea pigs, providing ample options for your experiment.
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