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: 0.1ug/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: 5-15ug/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™-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 and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store 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.
Immunogen
Recombinant protein corresponding to Met1-Glu118 from mouse DPPA5, expressed in E. coli.
Form
Supplied as a lyophilized powder in PBS, 5% trehalose. Reconstitute with 500ul sterile PBS.
Purity
Purified by immunoaffinity chromatography.
Specificity
Recognizes mouse DPPA5.