G25脱盐柱使用指南 (2)VIP

1、整理课件1Desalting column protocolMr. Yu Wang整理课件2 Matrix Sephadex G-25 Fine, cross-linked dextran Bed volume 53 ml Bed height 100 mm i.d. 26 mm separation of high (Mr 5 000) from low molecular weight substances (Mr 700) from smaller molecules (Mr 30 000 from molecules Mr 1 500 such as labeled protein o

2、r DNA from unconjugated dyes. The medium is often used to remove small nucleotides from longer chain nucleic acids.Sephadex G-25 is recommended for the majority of group separations involving globular proteins. These media are excellent for removing salt and other small contaminants away from molecu

3、les that are greater than Mr 5000.整理课件9sample volumes of up to 30% of the total volume of the desalting column can be processed. not exceed approximately 70 mg/ml。整理课件10Column: HiTrap Desalting, 1 5 ml, 3 5 ml, 5 5 mlSample: 2 mg/ml BSA in 50 mM sodium phosphate, 0.5 M sodium chloride, pH 7.0Sample

4、volume: 28% Vt (1.4, 4.3 and 7.1 ml respectively)Buffer: 50 mM sodium phosphate, 0.15 M sodium chloride, pH 7.0Flow rate: 5 ml/min整理课件11StorageStore unused media 4C to 30C in 20% ethanol. Do not freeze.Wash used media with 2 column volumes of distilled water followed by 2 column volumes of 20% ethan

5、ol. Store at 4C to 30C.Alternatively, wash with 2 column volumes of distilled water followed by 2 column volumes 0.01 M NaOH. Sodium hydroxide solution is bacteriostatic, easily disposed of and does not shrink the medium.Degas the ethanol/water mixture thoroughly and use a low flow rate, checking the back pressure as the column equil