The peptides synthesized on a derivatized cellulose membrane were probed with sera from patients who were allergic to Jun a 1, and the experimental data were interpreted with a PD classification method.
Purified, soluble IgG subclasses 1 4, each immobilized onto a nitrocellulose membrane, were probed with EndoS and EndoS E235Q) followed by incubation with antibodies against EndoS.
Both halves of the membrane were probed separately with anti-HA or anti-NFAT5 antibodies respectively and, before developing them by ECL, were realigned following the markers of the sliced lane.
The membrane were probed with either a primary IgG antibody (clone 3E8) or primary chicken anti-protein A IgY antibody (clone SPA-27) overnight at 4°C with constant inversion.
Membrane were probed with phosphor-specific Rb (Ser), Phospho-specific ERK (T/Y), Phospho-specific S6 (S235/236), Rab 11 primary antibodies (Cell Signaling Technology, Denver, MA), followed by AlexaFluor® 680-conjugated secondary antibodies (Invitrogen, Carlsbad, CA).
The membrane were probed with either: rabbit polyclonal anti-Bim (Cat No 202000, EMD Biosciences, San Diego, CA, USA), mouse monoclonal anti-flag (M2, Sigma), anti-Hsp90 (SPA-830, Stressgen, Victoria, Canada), mouse monoclonal anti-actin (ab6276 Abcam, Cambridge, UK) or mouse monoclonal anti-p23 (JJ3, generously supplied by Dr. David O Toft, Mayo Clinic, Rochester, MN, USA).
The membrane was probed with anti-myc (1∶3000).
The membrane was probed with antibody to dinitrophenylhydrazone derivatives (Dako, Glostrup, Denmark).
The PVDF membrane was probed with a mouse anti-myc antibody (clone 9E10, Cell Signaling Technologies).
After thorough washing with buffer A, the membrane was probed for caspase-9.
The membrane was probed with NrdA, NrdB or SeqA polyclonal antibodies.
