Goat Anti-Developmental Pluripotency-Associated Protein 4 (DPPA4)
DPPA4 is a 33kD molecule that is expressed specifically in the early embryo, uncommitted cells of the germ line, and embryonic stem cells and is used as a marker of pluripotent cells. Mouse DPPA4 is 296 amino acids in length and contains an SAP (SAF-A/B, ACINUS, PIAS) putative DNA-binding domain. Two potential isoforms exist with deletions of amino acids 104-120 and 69-120, respectively. In the region used as an immunogen, mouse DPPA4 shows 57% amino acid sequence identity with human and 87% identity with rat DPPA4 proteins.
Applications
Suitable for use in ELISA, Western Blot and Immunocytochemistry. Other applications not tested.
Recommended Dilutions
Western Blot: 0.1-0.2ug/ml with the appropriate secondary reagents to detect mouse DPPA4. The detection limit for rmDPPA4 is approximately 1 ng/lane under non-reducing and reducing conditions. This antibody has not been optimized for Western blot detection of DPPA4 in cell lysates. Immunocytochemistry: 10ug/ml to detect mouse DPPA4 in mouse embryonic stem cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at room temperature and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at room temperature. After blocking, cells were incubated with diluted primary antibody overnight at 4° C followed by Rhodamine RedTM-coupled anti-goat IgG at room termperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. Direct ELISA: This antibody can be used at 0.5-1.0ug/ml with the appropriate secondary reagents to detect mouse DPPA4. The detection limit for rmDPPA4 is approximately 0.2 ng/well. Optimal dilutions should be determined by each laboratory for each application.
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
E. coli-derived rmDPPA4 (aa125-296)
Form
Supplied as a lyophilized powder from a 0.2um filtered solution in PBS with 5% trehalose. Reconstitute with sterile PBS, 40-50% glycerol. If 0.5ml is used, the concentration will be 0.2mg/ml.
Purity
Purified by immunoaffinity chromatography.
Specificity
Selected for its ability to recognize mouse DPPA4 in the applications listed. In direct ELISAs and Western Blots, this antibody shows less than 5% crossreactivity with rhDPPA4 and rmDPPA5.
