Membrane was stained with Ponceau (Sigma) to validate transfer for 10 minutes in gentle agitation then de-stained for 1 hour with MQ water. Membrane was blocked with Licor blocking buffer (LIC927-70001) for 1 hour, washed three times for 5 minutes with TBS-T and incubated with primary antibody BTK (D3H5) Rabbit mAb (CST; 8547 S) overnight (16 hours) at 4°C, washed three times for 5 minutes with TBS-T and incubated with primary antibody against β-actin (CST; 3700) for 1 hour at 25°C. Membrane was washed three times for 5 minutes with TBS-T and incubated with a fluorescent secondary antibodies Anti-Mouse IgG Atto 488 (Sigma; 62197) and Anti-Rabbit-IgG Atto 647N (Sigma; 40839) for 1 hour, then washed three times for 5 minutes with TBS-T, dried and immediately imaged and analysed using Licor odyssey CLx. Prism (GraphPad) software was used to calculate degradation levels, and we used second order polynomial fit to estimate DC50 and Dmax values.