A Western blot loading control estimates lane-to-lane sample differences. A reference protein, such as actin, must have stable abundance and detection responsive to amount. Equal saturated bands fail to establish equal loading. [ref]
Considerations when quantitating protein abundance by immunoblot by Alicia A. McDonough and others. McDonough and others, “Consider the Loading Controls”, Figure 4: sample doubling changes claudin detection but not the actin signal
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