TABLE 3

Protein electrophoresis and isoelectric focusing methods

LocusProtein or gene nameE.C. numberTissue usedMethodaBuffer systembStaining referencec
ACP2 Acid phosphatase 23.1.3.2LiverSGETC-1Shaw and Prasad (1970)
ACONC Aconitase C4.2.1.3LiverSGECAAPM 1:19Richardson et al. (1986)
AK1 Adenylate kinase 12.7.4.3RbcSGETC-2Richardson et al. (1986)
ALDOC Aldolase C4.1.2.13BrainSGECAAPM 1:9Richardson et al. (1986)
AT3 Antithrombin 3SerumdPAGIEFIEF-1Aivaliotis et al. (1995)
C6 Complement component 6SerumdAGIEFIEF-2Aivaliotis et al. (1995)
DIA1 Diaphorase 11.6.2.2RbcSGEPHOS-1Harris and Hopkinson (1976)
GAPD Glyceraldehyde-3-phosphate dehydrogenase1.2.1.12RbcCAGETMWilliams (1964)
GPT Glutamate-pyruvate transaminase2.6.1.2RbcCAGEPHOS-2Richardson et al. (1986)
ME1 Malic enzyme 11.1.1.40BrainSGECAAPM 1:9Selander et al. (1971)
LDHB Lactate dehydrogenase B1.1.1.27RbcSGETC-2Selander et al. (1971)
PI Protease inhibitorSerumPAGIEFIEF-3van Oorschot et al. (1992b)
TF TransferrinSerumPAGETBThis study
  • a Methods: SGE, starch gel electrophoresis; PAGE, polyacrylamide gel electrophoresis; CAGE, cellulose acetate gel electrophoresis; AGIEF, agarose gel isoelectric focusing; PAGIEF, polyacrylamide gel isoelectric focusing. See text for details.

  • b Buffer systems: TC-1: 0.155 m Tris-0.043 m citrate, pH 7.0; system I of Shaw and Prasad (1970). TG2: 0.687 m Tris-0.157 m citrate, pH 8.0; system 5 of Selander et al. (1971). CAAPM 1:19: citric acid-aminopropyl morpholine, pH 6.0, modified from Clayton and Tretiak (1972); electrode buffer, 0.04 m citric acid, titrated to pH 6.0 with N-(3 aminopropyl)-morpholine; gel buffer, 1:19 dilution of electrode buffer. CAAPM 1:9: citric acid-aminopropyl morpholine, pH 6.0, modified from Clayton and Tretiak (1972); electrode buffer, 0.04 m citric acid, titrated to pH 6.0 with N-(3 aminopropyl)-morpholine; gel buffer, 1:9 dilution of electrode buffer. PHOS-1: 0.2 m phosphate, pH 5.8; system XIX of Shaw and Prasad (1970). PHOS-2: 0.2 m phosphate, pH 7.0; system B of Richardson et al. (1986). TB: modified gradient gel system of Gahne et al. (1977) using 0.187 m Tris-sulfate running gel, pH 9.0, and single 0.125 m Tris-HCl stacking gel, pH 6.8. TM: 0.05 m Tris-0.020 m maleic acid, pH 7.8; system C of Richardson et al. (1986). LEF-1: modified from Pollitt and Bell (1983) and Bell et al. (1992); equal parts of pH 5–6 (Pharmacia, Peapack, NJ) and pH 3–10 (SERVA Electrophoresis GmbH, Heidelberg, Germany) ampholines; anolyte, 0.04 m glutamic acid; catholyte, 0.1 m NaOH. IEF-2: modified from van Oorschot et al. (1993); 7:7:4 ratio of pH 4–6.5 (Pharmacia) :pH 5–6 (SERVA) :pH 3–10 (SERVA) ampholines; anolyte, 0.04 m glutamic acid; catholyte, 0.1 m NaOH. IEF-3: modified from van Oorschot et al. (1992b); 2:2:1 ratio of pH 4–5 (SERVA) :pH 4–5.5 (SERVA) :pH 3–10 (SERVA) ampholines; anolyte, 0.3 m phosphoric acid; catholyte, 1.0 m NaOH.

  • c Visualization of protein polymorphisms was accomplished by methods modified from the published or unpublished sources listed. Details of modified and unpublished procedures are available online at the Genetics supplemental information website: http://www.genetics.org/supplemental.