Anti-Developmental Pluripotency-Associated Protein 5 (DPPA5, ECAT2, Esg-1)

DPPA5, also known as Esg-1 and ECAT2, is a 14kD RNA-binding protein expressed in early embryos, primordial germ cells, and embryonic stem cells and is used as a marker of the pluripotent state. Mouse DPPA5 is 118 amino acids in length and shows 63% amino acid identity with human and 81% amino acid identity with rat DPPA5 proteins. Applications: Suitable for use in Western Blot, Immunocytochemistry and Direct ELISA. Other applications not tested. Recommended Dilutions: Western Blot: 0.1-0.2ug/ml with the appropriate secondary reagents to detect mouse DPPA5. The detection limit for rmDPPA5 is ~5ng/lane under non-reducing and reducing conditions. In this format, D3221-94T shows ~25% crossreactivity with rhDPPA5 and ~10% crossreactivity with rmDPPA4. D3221-94T has not been optimized for Western Blot detection of DPPA5 in cell lysates., Immunocytochemistry: 10ug/ml detects mouse DPPA5 in mouse embryonic stem cells. Cells were fixed with PBS, 4% paraformaldehyde for 20 minutes at RT and blocked with PBS, 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at RT. After blocking, cells were incubated with diluted D3221-94T overnight at 4°C followed by Rhodamine Red(TM)-coupled anti-goat IgG at RT in the dark for 1 hour. Between each step, cells were washed with PBS, 0.1% BSA. , Direct ELISA: 0.5-1.0ug/ml with the appropriate secondary reagents to detect mouse DPPA5. The detection limit for rmDPPA5 is ~0.3ng/well. , Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.