Electrophoresis in Practice (A Guide to Methods and Applications of DNA and Protein Separations) || Method 6: PAGIEF in Rehydrated Gels

✍ Scribed by Westermeier, Reiner

• Publisher: Wiley-VCH Verlag GmbH & Co. KGaA
• Year: 2005
• Tongue: German
• Weight: 278 KB
• Edition: 4
• Category: Article
• ISBN: 3527311815
• DOI: 10.1002/3527603468.ch13

✦ Synopsis

Method 6: PAGIEF in rehydrated gels 1 Sample preparation . Marker proteins pI 4.7 to 10.6 or . Marker proteins pI 5.5 to 10.7 + 100 lL of distilled water. . Meat extracts from pork, rabbit, veal and beef frozen in portions. Dilute before use: 100 lL of meat extract + 300 lL of double-distilled water. . Other samples: Set the protein concentration around 1 to 3 mg/mL. Dilute with distilled water. The salt concentration should not exceed 50 mmol/L.

It might be necessary to desalt with a NAP-10 column: apply 1 mL sample solution -use 1.5 mL of eluent.

2 Stock solutions

Acrylamide, Bis (T = 30 %, C = 3 %):

29.1 g of acrylamide + 0.9 g of Bis, make up to 100 mL with distilled water or PrePAG Mix (29.1 : 0.9), reconstitute with 100 mL of distilled water.

& Caution!

Acrylamide and Bis are toxic in the monomeric form. Avoid skin contact and do not pipette by mouth.