| 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. |
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| Catalog # | D3221-94S |
| Applications | Western Blot: This antibody can be used at 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: This antibody has been used at a concentration of 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 powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and 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. |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Goat |
| Source | Mouse |
| Concentration | As reported |
| 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 mouse DPPA4 affinity chromatography. |
| Immunogen | E. coli-derived rmDPPA4 (aa125-296) |
| 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. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
