Anti-Oct-4 (POU5F1)

Octamer-4 (Oct-4), a member of the POU family of transcription factors, has been demonstrated to be vital for the formation of self-renewing pluripotent stem cells. During embryogenesis, expression of Oct-4 is limited to pluripotent cells of the inner cell mass (ICM) that contribute to the formation of all fetal cell types. Applications: Suitable for use in Western Blot, Immunohistochemistry and Immunocytochemistry. Other applications not tested. Recommended Dilutions: Western Blot: 1:200-1:1000 using ECL, Immunohistochemistry: 1:200-1:1000, Immunocytochemistry: 1:200-1:1000, Optimal dilutions to be determined by the researcher. Immunohistochemistry Protocol: 1. Dissect tissues and freeze on dry ice., 2. Cut on a cryostat, 10um slices., 3. Dry slides at RT for 30-90 min., 4. Fix with cold Acetone:Methanol (50:50) 2 min. Either process further or store at -20ºC. Alternative is fixation with 4% PFA and treatment with 0.05% trypsin solution (incubation 10 min. at 37ºC, followed by 3 washes with PBS)., 5. Let air dry., 6. PBS 5 min., 7. Incubate in 50mM ammonium chloride 30 min., 8. PBS 5 min., 9. Blocking serum 30-45 min., 10. Primary antibody (O5403) 90 min., 11. Wash 3X for 5 min., 12. Secondary fluorescent conjug. antibody 30 min., 13. Wash 3X for 5 min., 14. Mount. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.