Recombinant Anti-RON antibody [EP1132Y] (ab52927)

Thank you for contacting Abcam regarding ab52927. I have confirmed that this antibody is an IgG1 isotype. Below, please find the IP protocol used: 1. Mix 200–300 μl of non-denatured cell lysate with appropriate dilutions of primary antibody 2. Incubate overnight at 4 °C with gentle rocking. 3. Add 25 μl of Protein G beads (50% bead slurry) and incubate 2 hours at 4 °C with gentle rocking. 4. Microcentrifuge for 30 seconds and discard the supernatant. Wash pellet 3 times with cold PBS. 5. Resuspend the pellet with 20 μl of 2x SDS reducing sample buffer containing β-mercaptoethanol (5%, v/v) and heat the sample at 95–100 °C for 3 minutes. 6. Briefly centrifuge the sample. 7. Load 16 μl of sample on to a SDS-PAGE gel or freeze the sample for later use (reboiled for 3 minutes prior loading onto the SDS-PAGE gel). 8. Perform Western blotting analysis using HRP-conjugated anti-rabbit IgG as secondary antibody  I hope this information is helpful.  Please do not hesitate to contact us if you have any additional questions.