For non-radioactive detection of receptor/ligand interaction.
This single vector system is the third generation AP-TAG technology. A secreted ligand or soluble receptor can be fused with
secreted alkaline phosphatase (AP) at either its N- or C-terminus to produce an "AP-body". The resulting AP fusion protein can be
expressed as a secreted protein and used directly as highly sensitive affinity agents much like an antibody. The epitope tags (6xHis and myc)
allow easy purification, detection and interaction assays (IP) of the AP-fusion proteins. Other improved features are listed below.
| | Cat. No. |
|---|
| AP-TAG Kit B - Academic | Q202 |
| AP-TAG Kit B - Industry | Q202P |
|
NOTE: pAPtag-5 plasmid vector can be transformed or
propagated in GH Competent cells (Cat. No. L301).
The L(left)-AP5 and R(right)-AP primers flanking the cloning sites of pAPtag-5 are used to check for the presence and size of DNA insert
cloned into the vector and for sequence verification of the Ligand-AP or soluble receptor-AP fusion constructs (N-terminal of AP only).
For C-terminal cloning, the L-AP5C and R-AP5C primers (not included in the kit) can be used (see below).
This kit is shipped on dry ice via overnight delivery.
References:
- Flanagan, J.G., et al. (2000). Alkaline phosphatase fusions of ligands or receptors as in situ probes for staining of cells, tissues and embryos. Methods in Enzymology 327, 17-35.
- Flanagan, J.G., and Cheng, H.-J. (2000). Alkaline phosphatase fusion proteins for molecular characterization and cloning of ligands and receptors. Methods in Enzymology 327, 198-210.
- US patents 5,554,499 and 5,801,000.
See AP-TAG Citation List for more.
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