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AP-TAG Kit B

$1,030.00
SKU:
Q202
Quantity:
Description
Specifications
Ordering
Documents

Product Description

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.

  AP-TAG Kit B  
  pAPtag-5 (20 µg) 40 µL
  L-AP5 primer (2 µM) 100 µL
  R-AP primer (2 µM) 100 µL
  Colony lysis buffer 2 x 1 mL
 Features pAPtag-5
 Size (kb) 5.8
 AP fusion type

Ligand-AP or AP-Ligand

(Receptor-AP or AP-Receptor)

 Cloning sites

Sfi I, Hind III, Bgl II, BspE I

or Xho I, Eco47 III, Xba I, Apa I

 Promoter CMV
 SV40 Ori.
 Yes
 E. coli host
GH (no need for P3)
 Vector selection
Ampicillin
 Secretion signal
 From vector or insert

 AP negative control  

(secretes AP alone)

Yes

 Affinity tags

6XHis or myc

 Transfection marker

Zeocin (Invitrogen)

NOTE: pAPtag-5 plasmid vectors 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. A detailed step-by-step protocol is included.

 

Purchase of an AP-TAG Ligand-Receptor Interaction Detection System (covered by US patents 5,554,499 and 5,801,000) comes with a limited, single-user and non-transferable sublicense for use in research applications only. If you plan to use AP-TAG technology to conduct drug screening or other commercial applications, a separate sublicense is required. Please contact GenHunter for details regarding the terms and conditions or any other questions. 

 

References:

  1. 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.
  2. 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.
  3. US patents 5,554,499 and 5,801,000
Individual components for the AP-TAG Kit B sold separately
 Description Volume

Cat. No.

(Academic)

Cat. No.

(Industry)

 pAPtag-5 (20 µg) 40 µL QV5 QV5P 
 L-AP5 primer (2 µM) 100 µL  Q212 Q212
 R-AP primer (2 µM) 100 µL  Q211 Q211 
 L-AP5C primer (2 µM) 100 µL Q215 Q215
 R-AP5C primer (2 µM) 100 µL  Q216 Q216
 Colony Lysis Buffer 5 mL L102 L102 
  AP-TAG Kit B  
  pAPtag-5 (20 µg) 40 µL
  L-AP5 primer (2 µM) 100 µL
  R-AP primer (2 µM) 100 µL
  Colony lysis buffer 2 x 1 mL
 Features pAPtag-5
 Size (kb) 5.8
 AP fusion type

Ligand-AP or AP-Ligand

(Receptor-AP or AP-Receptor)

 Cloning sites

Sfi I, Hind III, Bgl II, BspE I

or Xho I, Eco47 III, Xba I, Apa I

 Promoter CMV
 SV40 Ori.
 Yes
 E. coli host
GH (no need for P3)
 Vector selection
Ampicillin
 Secretion signal
 From vector or insert

 AP negative control  

(secretes AP alone)

Yes

 Affinity tags

6XHis or myc

 Transfection marker

Zeocin (Invitrogen)

Documents

AP-TAGbrochure-pdf.jpg"

AP-TAG Kit B - Cat. No. Q202 (for Academic/Non-profit institutions)

AP-TAG Kit B - Cat. No. Q202P (for Industry)

 

Purchase of an AP-TAG Ligand-Receptor Interaction Detection System (covered by US patents 5,554,499 and 5,801,000) comes with a limited, single-user and non-transferable sublicense for use in research applications only. If you plan to use AP-TAG technology to conduct drug screening or other commercial applications, a separate sublicense is required. Please contact GenHunter for details regarding the terms and conditions or any other questions. 

Individual components for the AP-TAG Kit B sold separately
 Description Volume

Cat. No.

(Academic)

Cat. No.

(Industry)

 pAPtag-5 (20 µg) 40 µL QV5 QV5P 
 L-AP5 primer (2 µM) 100 µL  Q212 Q212
 R-AP primer (2 µM) 100 µL  Q211 Q211 
 L-AP5C primer (2 µM) 100 µL Q215 Q215
 R-AP5C primer (2 µM) 100 µL  Q216 Q216
 Colony Lysis Buffer 5 mL L102 L102 

 

We also offer the AP-TAG Kit A (Q201) with pAPtag-2 and pAPtag-4 vectors.