文档介绍：试验室层析柱装柱技术介绍
AKTA实验室层析柱装柱技术介绍
第1页
Column packing
好柱效是层析成功关键
不一样种类凝胶、不一样层析柱有不一样装柱方法
柱效测定方法
层析柱维护
AKTA实验室层析
稳定和平衡凝胶柱床
检验柱效，必要时重装
Column packing
AKTA实验室层析柱装柱技术介绍
第10页
用空柱子须留心.
使用调整器(flow adaptor) - 保护柱床同时更可提升分辨率，节约凝胶
柱子物料须生物兼容，并有优良化学抗性
加上装柱器(packing reservoir)，可一次把凝胶装满柱子，大大提升柱效，尤其是凝胶过滤柱。
柱子过滤网孔径应该是凝胶颗粒1/3, 普通10mm孔径就够用，如用SOURCE15介质装C柱和XK柱，须换上5mm网
2021/10/4
AKTA实验室层析柱装柱技术介绍
第11页
Column packing
Sepharose FF based gel: Sepharose Q, DEAE, SP, Phenyl

1. the 75 % slurry just before packing.
2. Ideally, Fast Flow media are packed at a con stant pressure not exceeding 3 bar ( MPa) for XK columns.
3. If the packing equipment does not include a pressure gauge, use a packing flow rate of at least 400 cm/h (15 cm bed height, 25℃).
If recommended pressure or flow rate can not be obrained, use the maximum flow the pump can deliver, this should also give a easonably well-packed gel.
Do not exceed 75 % of the packing flow rate in subsequent chromatographic procedures.
AKTA实验室层析柱装柱技术介绍
第12页
Column packing
Sepharose 4 Fast Flow
50-75% slurry just before packing.
110 cm/h flow rate and the pressure should not be increased beyond bar
3-5mm blow the settled bed was made
AKTA实验室层析柱装柱技术介绍
第13页
Column packing
Sepharcyl S HR
Flow rates given below. Pack the gel in STEP 1 for 2 hours or until the gel has reached a constant height, then increase the flow rate to the value listed for STEP 2 and pack for 60 minutes.
Packing flow rates (ml/h):
Column STEP 1 STEP 2
XK 16/70 60 110
XK 16/100 60 100
XK 26/70 150 300
XK 26/100 150 270
XK 50/100 500 800
AKTA实验室层析柱装柱技术介绍
第14页
Column packing
Superdex 75,200 pg
50% slurry
1 step: 20cm/h flow rate, 2 hour
2 step: step the pressure is held at 3 bar, 1hour
AKTA实验室层析柱装柱技术介绍
第15页
Column packing
Sephadex G-25
50% slurry
400 cm/h flow rate
3mm blow th