H94N-apoCA-tagRFP Biosensor Protein

H94N-apoCA-tagRFP Biosensor Protein is a fluorescence-based biosensor with high sensitivity (KD = 0.22 nM) for measuring zinc.

Together with 4-(dimethylaminophenyl)benzoxazolyl sulfonamide, one can measure free zinc in aqueous solution at concentrations in the picomolar range (PMID 22430627, PMID 19152866, PMID: 17163650). It responds rapidly (see figure) to changes in free zinc at low levels which makes it useful for following rapid changes in free zinc concentration. It is one of our excitation ratiometric sensors, which makes accurate, reproducible calibration simple and minimizes most artifacts.

Read plog post from Dr. Richard Thompson at Pokegama Technologies, Improving Zinc Ion Quantitation with Unique Biosensor Proteins and Buffers.

Catalog Number Product DataSheet Size AVAILABILITY Price Qty
KPG007
H94N-apoCA-tagRFP Biosensor Protein
100ug In stock
Regular Price:$280.00
On Sale:
Specifications

Product Type: Protein
Name: H94N-apoCA-tagRFP biosensor protein
Extinction Coefficient: ε555 nm = 100,001
Molecular Weight: 60,000 Da
Variant MPN: T-0010
Format: Solution in pH 7.5 buffer
Purity: >95%
Tested Applications: Measuring free zinc ion concentrations
Comments: Fluorescent-labeled zinc free apoprotein; fluorescent label excitation maximum 555 nm, preferred emission wavelength 610 nm
Storage: +4C
Shipped: Cold packs

Data
H94N_Data
Comments
The sensor has extremely high affinity for Zn2+ (KD Å 0.22 nM), as well as Cu2+, Hg2+, Cd2+, Co2+, and Ni2+, with affinities mostly in the picomolar to nanomolar range. Thus the sensor should only be dissolved in metal-free buffers (or our metal ion buffers with known free zinc concentration), and should not touch metal or glass; most plastic (including natural pipette tips) is satisfactory in this regard. Ca2+ at 10 mM and Mg2+ at 50 mM do not interfere. For further details see our review Bozym 2008 PMID 19152866.
References
  1. T. K. Hurst, D. Wang, R.B. Thompson, C.A. Fierke, ÒCarbonic anhydrase II-based metal ion sensing: Advances and new perspectives,Ó Biochimica et Biophysica Acta (Proteins and Proteomics)1804, 393-403 (2010). PMID: 19818877; NIHMSID 157045.
  2. R.A. Bozym, T. Hurst, N. Westerberg, A.V. Stoddard, C.A. Fierke, C.J. Frederickson, R.B. Thompson, ÒDetermination of zinc using carbonic anhydrase-based fluorescence biosensors,Ó in Methods in Enzymology: Fluorescence Spectroscopy Vol. 450 (L. Brand and M.L. Johnson, editors) New York: Elsevier, pp 279-301 (2008). PMID 19152866.
  3. R. A. Bozym, A. K. Stoddard, C. A. Fierke, and R. B. Thompson, ÒMeasuring picomolar exchangeable zinc in PC-12 cells using a ratiometric fluorescence biosensor,Ó ACS Chemical Biology1(2) 103 Ð 111 (2006) PMID: 17163650

If you publish research with this product, please let us know so we can cite your paper.

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