Name: CM-Sepharose FF Gravity Prepacked Column


English name: CM NUPharose Fast Flow, CM NUPharose FF


Catalog number: NRPB23L, NRPB23S (prepacked column)


Specifications: 20 ml, 100 ml, 1L, 1 ml pre-packed column, 5 ml pre-packed column, special specifications customized


Transportation: 2 ~ 25℃, normal pressure, avoid light


Storage: 20% ethanol, 2 ~ 25℃


Shelf life: 5 years






Related introduction:


CM-Sepharose FF is a chromatographic separation medium with weak acid cation groups formed by bonding carboxymethyl groups on agarose gel microspheres. The product retains the excellent hydrophilicity of agarose and the large grid structure, and has good compatibility with biologically active macromolecules. It has the characteristics of high ion exchange capacity, low non-specific adsorption, and fast flow rate. It is widely used in Laboratory-scale preparation of biological macromolecules such as proteins, nucleic acids, peptides and polysaccharides, and industrial preparation of biopharmaceuticals and bioengineering.






Technical index:


Exterior


Milky white translucent gelatinous microspheres


Matrix


6% cross-linked agarose


Ion exchange type


Weak acid cationic group


Ligand


-O-CH2COO-


Ligand Density


100 ~ 180μmol/ml


Protein load


About 50 mg lysozyme/ml


Maximum flow rate


800 cm/h


Recommended flow rate


100 cm/h


Particle size range


60 ~ 180μm


Back pressure resistant


0.3 MPa


Operating temperature


4 ~ 40℃


Heat resistant


Sterilize in pH 7 water at 120°C for 30 min


pH stability


4 ~ 13 (long time); 2 ~ 14 (short time)


Chemical stability


Stable in the following solutions: commonly used aqueous buffer; 1 mol/L sodium hydroxide; 8 mol/L urea; 6 mol/L guanidine hydrochloride; 70% ethanol.






Instructions:


　　1. Column loading


　　1.1 Prepare initial buffer (equilibrium solution) and elution buffer according to the nature of the separation target.


　　1.2 Drain the gel and wash it twice with distilled water to remove the stored ethanol. Use the initial buffer (according to the ratio of gel:buffer=3:1) to homogenize and degas.


　　1.3 Fix the column vertically, moisten the bottom end with water or buffer and keep the liquid level for a period of time.


　　1.4 Use a glass rod to guide the homogenate and pour it into the column along the inner wall of the column at one time to allow the gel to settle freely in the column.


1.5 Connect the movable column head at the top of the column, turn on the peristaltic pump, let the buffer flow through 5 column volumes at the operating flow rate during use, and then use 1.5 times the operating flow rate to flow through 5 column volumes, adjust the adapter column head to make it as close as possible to the glue surface , And finally equilibrate the column with 2 to 3 column volumes of buffer.






Notice:


1) No air bubbles can be introduced during all operations to ensure the uniformity of glue filling.


2) If 1.2 is done unconditionally, there are bubbles in the packing layer, and the column can be loaded twice to remove the stored ethanol and bubbles.


3) The solution prepared by reagents such as ethanol needs to be degassed.


　　2. Balance


　　 Equilibrate the column with the equilibration buffer at the operating flow rate and observe the changes in the detector until the conductivity, pH and other parameters remain unchanged.


　　3. Sample loading


Switch the switching valve to load the sample. The sample amount is selected according to the nature of the sample and the amount of chromatographic medium. Linear experiments can also be performed to find the best sample amount; sample pretreatment: desalination, replacement of buffer, clarification and filtration ( 0.45, 0.22 μm) and so on.


　　4. Rinse


　　 Wash the loaded chromatography column with 2 to 3 column volumes of equilibration buffer and observe the changes of the detector until the conductivity, pH and other parameters remain unchanged, at which time the unexchanged components are washed out.


　　 5. Elution


　　Ion exchange column elution can be performed by constant elution, gradient elution or step elution. Generally, gradient elution with increasing salt concentration is recommended.


　　 6. Regeneration


　　Use a high salt concentration buffer (containing 1 ~ 2mol/L NaCl) to flush 3 ~ 5 column volumes at the operating flow rate, and then wash with a balance solution to equilibrate 3 ~ 5 column volumes.


　　 If there are inactivated proteins or lipids that cannot be washed off during regeneration, they can be removed by cleaning-in-place (CIP).


　　7. Cleaning-in-place (CIP)


For strong binding proteins or lipids, the following process can be used for in-place cleaning: 1 mol/L NaOH for 2 column volumes, 8 mol/L urea for 2 column volumes, and 30% isopropanol for 2 column volumes , Wash 2 column volumes with equilibration buffer.


　　In-place cleaning can effectively remove impurities on the medium, and the reverse effect is better. If you need to remove endotoxin and pyrogen, you can clean it with 1 mol/L NaOH for 1 to 2 hours at a speed of 50 cm/h.

Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance

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Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance.The website pictures are from the Internet. The pictures are for reference only. Please take the real object as the standard. In case of infringement, please contact us to delete them immediately.