Abstract
Poly R‐480, an amphoteric dye of molecular weight 160 000,is the product of copolymerization reaction and can be expected to yield a polydisperse pattern in polyacrylamide gel isoelectric focusing (PAGIF). Such is indeed obtained in conventional PAGIF using commercial synthetic carrier ampholyte mixtures but not in buffer PAGIF, in which a few discrete dye bands are produced. Upon excision of single bands and segments of zones in buffer PAGIF and PAGIF in SCAMs, respectively, the entire original pattern is reestablished in each case, thereby confirming the artifactual nature of both the simple band pattern as well as the polydisperse patten. The simple pattern in buffer PAGIF prevails in the presence of 8 M urea and 8 mM CHAPS, while the polydisperse pattern obtained in SCAMs is resolved into multiple bands in the presence of either one or both of these agents. These observations suggest: a) hydrophobic and hydrogen bond dye‐dye and/or dye‐SCAM interactions produce the artifactual polydisperse pattern in PAGIF in SCAMs, b) electrostatic dye‐dye interactions are responsible for the artifactual simple band pattern in buffer EF, and c) dye‐SCAM electrostatic interactions produce the artifactual multiple band pattern in the SCAM‐urea‐CHAPS system. These data clearly underline the need for critical experimentation in the evaluation of PAGIF bands and band numbers.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 404-407 |
| Number of pages | 4 |
| Journal | Electrophoresis |
| Volume | 4 |
| Issue number | 6 |
| DOIs | |
| State | Published - 1983 |
| Externally published | Yes |
Fingerprint
ASJC Scopus subject areas
- Biochemistry
- Clinical Biochemistry
Cite this
Detection of binding artifacts in gel electrofocusing using a polymeric dye probe. / Cuono, Charles B.; Chapo, Giselle A.; Chrambach, Andreas; Hjelmeland, Leonard M.
In: Electrophoresis, Vol. 4, No. 6, 1983, p. 404-407.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Detection of binding artifacts in gel electrofocusing using a polymeric dye probe
AU - Cuono, Charles B.
AU - Chapo, Giselle A.
AU - Chrambach, Andreas
AU - Hjelmeland, Leonard M
PY - 1983
Y1 - 1983
N2 - Poly R‐480, an amphoteric dye of molecular weight 160 000,is the product of copolymerization reaction and can be expected to yield a polydisperse pattern in polyacrylamide gel isoelectric focusing (PAGIF). Such is indeed obtained in conventional PAGIF using commercial synthetic carrier ampholyte mixtures but not in buffer PAGIF, in which a few discrete dye bands are produced. Upon excision of single bands and segments of zones in buffer PAGIF and PAGIF in SCAMs, respectively, the entire original pattern is reestablished in each case, thereby confirming the artifactual nature of both the simple band pattern as well as the polydisperse patten. The simple pattern in buffer PAGIF prevails in the presence of 8 M urea and 8 mM CHAPS, while the polydisperse pattern obtained in SCAMs is resolved into multiple bands in the presence of either one or both of these agents. These observations suggest: a) hydrophobic and hydrogen bond dye‐dye and/or dye‐SCAM interactions produce the artifactual polydisperse pattern in PAGIF in SCAMs, b) electrostatic dye‐dye interactions are responsible for the artifactual simple band pattern in buffer EF, and c) dye‐SCAM electrostatic interactions produce the artifactual multiple band pattern in the SCAM‐urea‐CHAPS system. These data clearly underline the need for critical experimentation in the evaluation of PAGIF bands and band numbers.
AB - Poly R‐480, an amphoteric dye of molecular weight 160 000,is the product of copolymerization reaction and can be expected to yield a polydisperse pattern in polyacrylamide gel isoelectric focusing (PAGIF). Such is indeed obtained in conventional PAGIF using commercial synthetic carrier ampholyte mixtures but not in buffer PAGIF, in which a few discrete dye bands are produced. Upon excision of single bands and segments of zones in buffer PAGIF and PAGIF in SCAMs, respectively, the entire original pattern is reestablished in each case, thereby confirming the artifactual nature of both the simple band pattern as well as the polydisperse patten. The simple pattern in buffer PAGIF prevails in the presence of 8 M urea and 8 mM CHAPS, while the polydisperse pattern obtained in SCAMs is resolved into multiple bands in the presence of either one or both of these agents. These observations suggest: a) hydrophobic and hydrogen bond dye‐dye and/or dye‐SCAM interactions produce the artifactual polydisperse pattern in PAGIF in SCAMs, b) electrostatic dye‐dye interactions are responsible for the artifactual simple band pattern in buffer EF, and c) dye‐SCAM electrostatic interactions produce the artifactual multiple band pattern in the SCAM‐urea‐CHAPS system. These data clearly underline the need for critical experimentation in the evaluation of PAGIF bands and band numbers.
UR - http://www.scopus.com/inward/record.url?scp=84988141847&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84988141847&partnerID=8YFLogxK
U2 - 10.1002/elps.1150040607
DO - 10.1002/elps.1150040607
M3 - Article
AN - SCOPUS:84988141847
VL - 4
SP - 404
EP - 407
JO - Electrophoresis
JF - Electrophoresis
SN - 0173-0835
IS - 6
ER -