Anti-Progesterone Receptor抗体(ab63605)

Thank you for your enquiry. I have been in contact with the originator of ab63605 Progesterone Receptor antibody who have kindly provided the following ICC procedure. A. Soultions and Reagents 1. 10× PBS: To prepare 1L add 80g NaCl, 2g KCl, 2g KH2PO4 and 28.5g NaHPO4 to 1L water for injection. Adjust pH to 7.4. 2. 4% Polyoxymethylene: To prepare 100mL add 4g polyoxymethylene to 100mL 1×PBS. Adjust pH to 7.4. 3. 1×PBS/0.2% Triton X-100(PBS/Triton): To prepare 500mL add 1mL Triton X-100 to 500mL 1× PBS. 4. 1×PBS/3% BSA(PBS/BSA): To prepare 100mL add 3g BSA to 100mL 1× PBS. B. Preparation Fixation permeabilization 1. Rinse cells briefly in PBS. 2. Aspirate PBS, cover cells to a depth of 2-3mm about 200ul with 4% polyoxymethylene. 3. Allow cells to fix for 15 minutes at room temperature. 4. Aspirate fixative, rinse three times in PBS for 5 minutes each. 5. Aspirate PBS, cover cells to a depth of 2-3mm about 200ul with PBS/Triton for 5 minutes at room temperature. 6. Aspirate permeability agent rinse three times in PBS for 5 minutes each. C. Immunostaining 1. Gently add 200ul of primary antibody diluted in PBS/BSA to the 24 well plates each well. 2. Incubate 60 minutes at 37℃ or overnight at 4℃. 3. Aspirate diluted primary antibody, then rinse three times in PBS for 5 minutes each. 4. Incubate in fluorochrome-conjugate secondary antibody diluted in PBS/BSA to the 24 well plates 100ul each well for 30 minutes at room temperature in dark. 5. Aspirate diluted fluorochrome-conjugate secondary antibody, then rinse three times in PBS for 5 minutes each. 6. Test under fluorescence microscope. I hope this will be helpful to you. With regards to a blocking peptide for this antibody, I am sorry that for this particular product, there is no blocking peptide available. Should you have any further questions, please do not hesitate to contact us.