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.\n\nApplications: \nWestern 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. \nImmunocytochemistry: 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 degrees 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. \nDirect 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. \nOptimal dilutions should be determined by each laboratory for each application.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees 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.

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SPECIFICATIONS

Catalog Number

D3221-94S

Size

100ug

Applications

ELISA, ICC, WB

Hosts

Goat

Reactivities

Mouse

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.

P Type

Pab

Purity

Purified by mouse DPPA4 affinity chromatography.

Isotype

IgG

Additional Info

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.