The colloidal gold protein assay is a highly sensitive biochemicalassay for determining the total concentration of protein in a solution (~0.1 ng/μL to 200 ng/μL).[1] It was first described in 1987 by two groups who used commercially available "Aurodye" colloidal gold solutions.[2][3] Notably, the formulation of Aurodye changed between 1987 and 1990 such that it became incompatible with protein assays,[4] however vendors such as Bio-Rad[5] & Diversified Biotech[4] starting offering colloidal gold formulations that were suitable for protein assays. These products have since been discontinued and there are no vendors that currently explicitly sell colloidal gold for the assay, however detailed synthetic procedures were published to produce the ~17-40 nm gold nanoparticles that are suitable for the assay, along with modifications to increase the shelf stability of the colloidal gold & adapt the assay to microplate format & increase its sensitivity.[6] Gold nanoparticles in the ~17-40 nm size range that are presumably compatible with the assay are currently commercially available.[7]
The total protein concentration is readout by an increase in absorbance at 565 nm, which can then be measured using colorimetric techniques, including using microplate readers. Most common reagents, except thiols and SDS, are compatible with the assay.[1] An optimized formulation for the assay to maximize sensitivity in microplate format was described.[6]
Comparison to other assaysedit
While the colloidal gold assay is the most sensitive in-solution colorimetric protein assay, it may be equally sensitive or surpassed in sensitivity by fluorescent protein assays such as the CBQCA, FQ, NanoOrange, Quant-iT, and EZQ assays.[8]
^ abAhmed, Hafiz (2005). Principles and Reactions of Protein Extraction, Purification, and Characterization. CRC Press. p. 58. doi:10.1201/9780203507438. ISBN978-0-203-50743-8.
^Stoscheck, Christa M. (1987). "Protein assay sensitive at nanogram levels". Analytical Biochemistry. 160 (2). Elsevier BV: 301–305. doi:10.1016/0003-2697(87)90051-0. ISSN 0003-2697.
^Hunter, John B.; Hunter, Susan M. (1987). "Quantification of proteins in the low nanogram range by staining with the colloidal gold stain AuroDye". Analytical Biochemistry. 164 (2). Elsevier BV: 430–433. doi:10.1016/0003-2697(87)90515-x. ISSN 0003-2697.
^ abDeutscher, M.P. (1990). Guide to Protein Purification. Methods in enzymology. Academic Press. p. 64. ISBN978-0-12-182083-1. Retrieved 2023-04-09.
^Rad, Bio-. "Enhanced Colloidal Gold Total Protein Detection Kit - Instruction Manual" (PDF). Bulletin_9238.pdf. Bio-Rad. Retrieved 9 April 2023.
^ abCiesiolka, Thomas; Gabius, Hans-Joachim (1988). "An 8- to 10-fold enhancement in sensitivity for quantitation of proteins by modified application of colloidal gold". Analytical Biochemistry. 168 (2). Elsevier BV: 280–283. doi:10.1016/0003-2697(88)90319-3. ISSN 0003-2697.
^Burgess, R.R.; Deutscher, M.P. (2009). Guide to Protein Purification. ISSN. Elsevier Science. p. 80. ISBN978-0-08-092317-8. Retrieved 2023-04-09.
May 03, 2024
colloidal, gold, protein, assay, colloidal, gold, protein, assay, highly, sensitive, biochemical, assay, determining, total, concentration, protein, solution, first, described, 1987, groups, used, commercially, available, aurodye, colloidal, gold, solutions, n. The colloidal gold protein assay is a highly sensitive biochemical assay for determining the total concentration of protein in a solution 0 1 ng mL to 200 ng mL 1 It was first described in 1987 by two groups who used commercially available Aurodye colloidal gold solutions 2 3 Notably the formulation of Aurodye changed between 1987 and 1990 such that it became incompatible with protein assays 4 however vendors such as Bio Rad 5 amp Diversified Biotech 4 starting offering colloidal gold formulations that were suitable for protein assays These products have since been discontinued and there are no vendors that currently explicitly sell colloidal gold for the assay however detailed synthetic procedures were published to produce the 17 40 nm gold nanoparticles that are suitable for the assay along with modifications to increase the shelf stability of the colloidal gold amp adapt the assay to microplate format amp increase its sensitivity 6 Gold nanoparticles in the 17 40 nm size range that are presumably compatible with the assay are currently commercially available 7 Contents 1 Mechanism 2 Comparison to other assays 3 See also 4 ReferencesMechanism editThe total protein concentration is readout by an increase in absorbance at 565 nm which can then be measured using colorimetric techniques including using microplate readers Most common reagents except thiols and SDS are compatible with the assay 1 An optimized formulation for the assay to maximize sensitivity in microplate format was described 6 Comparison to other assays editWhile the colloidal gold assay is the most sensitive in solution colorimetric protein assay it may be equally sensitive or surpassed in sensitivity by fluorescent protein assays such as the CBQCA FQ NanoOrange Quant iT and EZQ assays 8 See also editColloidal gold Bradford assay BCA assayReferences edit a b Ahmed Hafiz 2005 Principles and Reactions of Protein Extraction Purification and Characterization CRC Press p 58 doi 10 1201 9780203507438 ISBN 978 0 203 50743 8 Stoscheck Christa M 1987 Protein assay sensitive at nanogram levels Analytical Biochemistry 160 2 Elsevier BV 301 305 doi 10 1016 0003 2697 87 90051 0 ISSN 0003 2697 Hunter John B Hunter Susan M 1987 Quantification of proteins in the low nanogram range by staining with the colloidal gold stain AuroDye Analytical Biochemistry 164 2 Elsevier BV 430 433 doi 10 1016 0003 2697 87 90515 x ISSN 0003 2697 a b Deutscher M P 1990 Guide to Protein Purification Methods in enzymology Academic Press p 64 ISBN 978 0 12 182083 1 Retrieved 2023 04 09 Rad Bio Enhanced Colloidal Gold Total Protein Detection Kit Instruction Manual PDF Bulletin 9238 pdf Bio Rad Retrieved 9 April 2023 a b Ciesiolka Thomas Gabius Hans Joachim 1988 An 8 to 10 fold enhancement in sensitivity for quantitation of proteins by modified application of colloidal gold Analytical Biochemistry 168 2 Elsevier BV 280 283 doi 10 1016 0003 2697 88 90319 3 ISSN 0003 2697 Spherical Gold Nanoparticles Nanopartz Retrieved 2023 04 09 Burgess R R Deutscher M P 2009 Guide to Protein Purification ISSN Elsevier Science p 80 ISBN 978 0 08 092317 8 Retrieved 2023 04 09 Retrieved from https en wikipedia org w index php title Colloidal gold protein assay amp oldid 1221075005, wikipedia, wiki, book, books, library,
