Example 3
Mouse monoclonal antibodies were purchased from Invitrogen (CUL1-2H4C9: #32-2400), Sigma (FLAG-M2: #F3165, β-actin: #A-1978), Covance (HA-16612: #MMS-101P) and Santa Cruz (CD147 8D6: #sc-21746): Rabbit polyclonal antibodies were from Millipore (MCT1: #AB-3538P), Bethyl (DDB1: #A300-462A, CUL4A: #A300-739A), Sigma (FLAG, #F7425), Cell Signaling (α/β-tubulin: #2148, IKZF1: #5443, IKZF3: #12720), Proteintech (CUL4B: #12916-1-AP) and Santa Cruz (HA Y-11: #sc-805. A polyclonal antibody against CRBN was generated by immunizing rabbits with a mixture of two peptides containing amino acids 1-19 and 424-437 of human CRBN (MAGEDQQDAAHNMGNHLPC (SEQ. ID NO: 1) and CPTIDPTEDEISPDK (SEQ ID NO: 2), respectively). Secondary antibodies (anti-rabbit IgG, anti-mouse IgG or protein-A coupled with horseradish peroxidase) were from GE Healthcare. Secondary antibodies (anti-rabbit IgG and anti-mouse IgG) coupled to Alexa Fluor 594 or Alexa Fluor 488 for immunofluorescence and flow cytometry were from Life technologies.