TRIS Buffer for the Demonstration of Haemoglobin A₂by Paper Electrophoresis

A new haemoglobin J from Turkey ‐ HB Ankara (β 10 (A7) Ala → Asp).

“…On paper electrophoresis at pH 8.9, the patient's haemolysate showed three fractions, namely [ 1 ] haemolysate into 3% Hb A2, 55% Hb A + Aa and 42% Hb J + Ja. Even after further purification by paper electrophoresis the Hb J fraction was seen, on fingerprinting, to still contain some Hb Aa.…”

“…Haemoglobin (Hb) electrophoresis was performed on paper in Tris buffer at pH 8.9 [1] and on starch gel at pH 8.6 [2]. Haemolysates were fractionated using column chromatography as described by Huisman and Dozy [3] ; the resulting haemoglobin fractions were concentrated by ultrafiltration at 4°C and purified by paper electrophoresis [1 ], Globin was prepared from purified haemoglobin by acid-acetone precipitation at -20°C [4], washed three times with cold acetone and dried in vacuo over P2Os.…”

“…Haemolysates were fractionated using column chromatography as described by Huisman and Dozy [3] ; the resulting haemoglobin fractions were concentrated by ultrafiltration at 4°C and purified by paper electrophoresis [1 ], Globin was prepared from purified haemoglobin by acid-acetone precipitation at -20°C [4], washed three times with cold acetone and dried in vacuo over P2Os. Digestion of the total globin with trypsin and analysis of that portion of the digest which was soluble at pH 6.4, by fingerprinting, was carried out as previously described [5].…”

Cellulose Acetate Membranes for the Electrophoretic Demonstration of Hemoglobin A₂

“…Goldberg (B,9) has reported excellent separation of A2 with starch-gel electrophoresis, employing dis continuous buffer systems. Craddock-Watson et al (10) reported the separation of A2 hemoglobin on filter paper employing TRIS buffer, but this method requires prolonged electrophoresis and does not lend itself to quantification.…”

Haemoglobin Stanleyville II (<i>α</i> 78 [EF 7] Asn → Lys) Found in France

“…globin fraction was concentrated and purified by repeated electrophoresis on filter paper in Tris-EDTA-borate buffer at pH 8.9 [6], Globin was prepared by precipi tation (1.5°/o v/v of concentrated HC1 in acetone) at 0 °C [7] and washed 3 times in acetone at 0 °C.…”