Love pure pie . . . The laboratory column encyclodeda came.

has been loved by many teachers since its launch. Whenever you have any questions about the purification of biomes, send an email to our exclusive email address akta.club@ge.com and an expert will answer your questions within 48 hours. Hung Hom frequently pass, in recent times teachers on the problem of loading columns more, in this article, we will introduce some of the key points of laboratory column loading, in order to win readers: 1, the choice of laboratory empty columns empty columns of the material has plastic, glass and metal, materials are different, empty columns can withstand the greatest pressure and application range is also different. GE offers empty columns made of glass for the separation and purification of a wide range of biomass molecules, in three main categories: XK columns, Tricon columns and HiScale columns.Air Column Classification 1, XK Column: Known as the "Small Prince of Medium and Low Pressure", it is mainly used for medium and low voltage laminate (0-5 bar) and is highly regarded for its excellent performanceadvantages: suitable for the vast majority of
fills
including Superdex and SuperHoseP, temperature controlled and pressure resistant.standard column height regulator AK adaptor provides flexibility in adjusting column height.flow regulator on the tightly sealed AK adaptor ensures smooth fluid flow and low dead volume (less than 0.1% of the total column volume) to minimize peak width.can be used when loading columns, or another empty column can be connected together with connector connector for mounting columns.2, Tricorn empty column: "High-resolution high-voltage preferred" for medium- and high-voltage layering systems for medium- and small-scale high-resolution analysis.
advantages:
high pressure resistance, up to 100 bar.use Valco connectors, directly connected to the HPLC/KTA liquid phase
chromatography
.buckle design prevents accidental rotation of the regulator and keeps the column bed stable.without pipes optimizes the column dead volume to near zero.can be filled with high-resolution fillers such as SOURCE, Superdex, Superose.
3, HiScale column: "High flow rate amplification first choice", the new pressure-resistant 20Bar empty column, more suitable for the next generation of high-flow fillers such as Capto and MabSelect series.
advantages:
20Bar pressure for modern high-flow
agar
sugar fillers such as Fast Flow Classic Fillers and Capto Mabselect, significantly reducing separation purification cycles.unique head design, better compatible with axial compression, to ensure higher column efficiency.the quickLock mechanism of the bar head to make the operation and cleaning of the bar easier and faster.with two bar-to-column heights provides high flexibility. Filler and empty column matching table, click to see the big picture, minutes to complete the column selection, take good not to send to 2, empty column preparation preparation steps: empty column screen needs to be regularly cleaned with 0.5 M NaOH ultrasonic cleaning, and with 20 % ethanol to remove air bubbles, internal contact must ensure tightening, each component after assembly with water to check air tightness. The empty column is fixed with a column clip, connected to the analyzer, to ensure horizontal and vertical, for the gel
filtration
filler filling, need to be checked with a level.
This, the preparation of filler slurry
repeated use of fillers: the need for regeneration of fillers, if the filler regeneration is still significant, after stirring part of the floc-shaped or pigmented calm, it is recommended to choose a new filler. New Filler: Filler storage solution is usually 20 % ethanol, column solution depending on the filler, may be aqueous solution, salt solution or
organic
solution, etc. The above has been exchanged into the column solution of the filler for homogeneous slurry, you can choose
soch
, the glass rod can be stirred lightly, cut not available mixers, to prevent the filler particles from breaking. The plasma concentration is between 50%-70 % and is too thin or too thick to affect the uniformity of the filling. The volume of the required filler slurry
4, refringing and pressing glue
a. Inject the lower end of the empty column into a column solution of about 2 cm height with a syringe. b. Then the filler is drained with a glass rod and poured into an empty column, the process is fast and gentle, preventing bubbles from creating. c. Replenish the column solution with a syringe to the top of the empty column, so that the liquid surface slightly protrudes on the upper end of the empty column, the upper column head screen is degassed after the tilt into the empty column, tightens the outer joint and the column head, sets the column front pressure (protects the empty column) and Delta pressure (protects the filler) after starting the laminate equipment to start pressing glue. Can be used by constant pressure or constant flow of the way of pressing glue (load column recommended flow rate or pressure can be seen in the filler manual), the difference between the two is as follows: d. after the glue surface no longer drops, mark the glue surface position, pause the purification instrument, loosen the column head, drop to the glue surface below about 3 mm, retightening can be.
, column effect detection . Detection method: is the index to detect the quality of the column, but also the key factor of the success or failure of the target molecular purification. Column efficiency is determined by theoretical tower height (HETP) and symmetry factor (As). The determination of column efficiency can be measured with 1 % acetone or 2 M NaCl, with UV 280 and conductivity respectively, the sample amount is 1 % - 2 % of the column volume, the flow rate in the determination of column effect can be found in the filler manual. Results Evaluation: Theoretical Tower Height (HETP): Ideally 1-2 filler particle sizes, usually 2-4 filler particle sizes are very good, 3-5 filler particle sizes are acceptable, and of course, depending on the SOP requirements in inherent production. (As): ideally 1, but 0.8-1.5 is also within acceptable range. out of the peak tail: indicates that the filler is not pressurized. flow out of the peak forward: indicating that the filler pressure is too tight. flow out of the peak: indicates that the density of fillers in the column is not equal.
. Expert Team Our 12 love pure experts dedicated to
protein purification
, or focus on blood products, or proficient in
antibodies
purification, or dedicated vaccine production, more all-powerful, hundred Xiaosheng ... There's always one for you! If you still have questions about the loading process, please send your questions to akta.club@ge.com and your questions will be answered within 48 hours

.