Thank you very much Adam. In our laboratory, we don't have automatic FPLC. Is it possible to use QAE-Sephadex column without requiring any other instrument?
Thank you very much Adam. In our laboratory, we don't have automatic FPLC. Is it possible to use QAE-Sephadex column without requiring any other instrument?
I am under the impression that DEAE-cellulose is no longer sold. You can substitute another ion-exchange resin such as QAE-Sephadex if you are planning open-top column chromatography, or QAE-Sepharose if you are planning to use an FPLC. Ion exchange chromatography is usually started at a low ionic strength, at which the proteins bind to the column. Then the column is eluted with a gradient of increasing ionic strength. Fractions are collected during the gradient elution. If the equipment is available, an automated FPLC system is great. However, simpler technologies can also be used.
I am under the impression that DEAE-cellulose is no longer sold. You can substitute another ion-exchange resin such as QAE-Sephadex if you are planning open-top column chromatography, or QAE-Sepharose if you are planning to use an FPLC. Ion exchange chromatography is usually started at a low ionic strength, at which the proteins bind to the column. Then the column is eluted with a gradient of increasing ionic strength. Fractions are collected during the gradient elution. If the equipment is available, an automated FPLC system is great. However, simpler technologies can also be used.
Hi Before setting up the column try to screen the ion exchange resins for best binding purpose. Screening can be done using less resin quantity and also gives u an idea about elution profile as which resin binding is weak or strong to proceed for column chromatography runs. As Dr. Adam has mentioned, FPLC is automated technology which saves time and gives best fold purity. If u do not have FPLC unit let me know will guide for gravity separations as well. Good luck
Hi Before setting up the column try to screen the ion exchange resins for best binding purpose. Screening can be done using less resin quantity and also gives u an idea about elution profile as which resin binding is weak or strong to proceed for column chromatography runs. As Dr. Adam has mentioned, FPLC is automated technology which saves time and gives best fold purity. If u do not have FPLC unit let me know will guide for gravity separations as well. Good luck
I am under the impression that DEAE-cellulose is no longer sold. You can substitute another ion-exchange resin such as QAE-Sephadex if you are planning open-top column chromatography, or QAE-Sepharose if you are planning to use an FPLC. Ion exchange chromatography is usually started at a low ionic strength, at which the proteins bind to the column. Then the column is eluted with a gradient of increasing ionic strength. Fractions are collected during the gradient elution. If the equipment is available, an automated FPLC system is great. However, simpler technologies can also be used.
I am under the impression that DEAE-cellulose is no longer sold. You can substitute another ion-exchange resin such as QAE-Sephadex if you are planning open-top column chromatography, or QAE-Sepharose if you are planning to use an FPLC. Ion exchange chromatography is usually started at a low ionic strength, at which the proteins bind to the column. Then the column is eluted with a gradient of increasing ionic strength. Fractions are collected during the gradient elution. If the equipment is available, an automated FPLC system is great. However, simpler technologies can also be used.
If you have a small peristaltic pump, a magnetic stirrer, a 2-chamber gradient former, and some appropriately sized flexible tubing, you can do ion-exchange chromatography without an FPLC. If you don't have a peristaltic pump, you can use gravity to elute the column. If you don't have a gradient former, you can use 2 beakers connected with tubing. You can also avoid making a continuous gradient and instead use a series of small step elutions of gradually increasing salt concentration. An automatic fraction collector is nice, but if you don't have that you can collect fractions by hand.
If you have a small peristaltic pump, a magnetic stirrer, a 2-chamber gradient former, and some appropriately sized flexible tubing, you can do ion-exchange chromatography without an FPLC. If you don't have a peristaltic pump, you can use gravity to elute the column. If you don't have a gradient former, you can use 2 beakers connected with tubing. You can also avoid making a continuous gradient and instead use a series of small step elutions of gradually increasing salt concentration. An automatic fraction collector is nice, but if you don't have that you can collect fractions by hand.
If you have a small peristaltic pump, a magnetic stirrer, a 2-chamber gradient former, and some appropriately sized flexible tubing, you can do ion-exchange chromatography without an FPLC. If you don't have a peristaltic pump, you can use gravity to elute the column. If you don't have a gradient former, you can use 2 beakers connected with tubing. You can also avoid making a continuous gradient and instead use a series of small step elutions of gradually increasing salt concentration. An automatic fraction collector is nice, but if you don't have that you can collect fractions by hand.
If you have a small peristaltic pump, a magnetic stirrer, a 2-chamber gradient former, and some appropriately sized flexible tubing, you can do ion-exchange chromatography without an FPLC. If you don't have a peristaltic pump, you can use gravity to elute the column. If you don't have a gradient former, you can use 2 beakers connected with tubing. You can also avoid making a continuous gradient and instead use a series of small step elutions of gradually increasing salt concentration. An automatic fraction collector is nice, but if you don't have that you can collect fractions by hand.
Thank you Dr Adam B Shapiro.
Thank you Dr Adam B Shapiro.
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Thank you very much Adam. In our laboratory, we don't have automatic FPLC. Is it possible to use QAE-Sephadex column without requiring any other instrument?
Thank you very much Adam. In our laboratory, we don't have automatic FPLC. Is it possible to use QAE-Sephadex column without requiring any other instrument?
More
VOTE
I am under the impression that DEAE-cellulose is no longer sold. You can substitute another ion-exchange resin such as QAE-Sephadex if you are planning open-top column chromatography, or QAE-Sepharose if you are planning to use an FPLC. Ion exchange chromatography is usually started at a low ionic strength, at which the proteins bind to the column. Then the column is eluted with a gradient of increasing ionic strength. Fractions are collected during the gradient elution. If the equipment is available, an automated FPLC system is great. However, simpler technologies can also be used.
I am under the impression that DEAE-cellulose is no longer sold. You can substitute another ion-exchange resin such as QAE-Sephadex if you are planning open-top column chromatography, or QAE-Sepharose if you are planning to use an FPLC. Ion exchange chromatography is usually started at a low ionic strength, at which the proteins bind to the column. Then the column is eluted with a gradient of increasing ionic strength. Fractions are collected during the gradient elution. If the equipment is available, an automated FPLC system is great. However, simpler technologies can also be used.
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Hi
Before setting up the column try to screen the ion exchange resins for best binding purpose. Screening can be done using less resin quantity and also gives u an idea about elution profile as which resin binding is weak or strong to proceed for column chromatography runs. As Dr. Adam has mentioned, FPLC is automated technology which saves time and gives best fold purity. If u do not have FPLC unit let me know will guide for gravity separations as well.
Good luck
Hi
Before setting up the column try to screen the ion exchange resins for best binding purpose. Screening can be done using less resin quantity and also gives u an idea about elution profile as which resin binding is weak or strong to proceed for column chromatography runs. As Dr. Adam has mentioned, FPLC is automated technology which saves time and gives best fold purity. If u do not have FPLC unit let me know will guide for gravity separations as well.
Good luck
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VOTE
Thank you Dr Adam B Shapiro.
Thank you Dr Adam B Shapiro.
More
VOTE
I am under the impression that DEAE-cellulose is no longer sold. You can substitute another ion-exchange resin such as QAE-Sephadex if you are planning open-top column chromatography, or QAE-Sepharose if you are planning to use an FPLC. Ion exchange chromatography is usually started at a low ionic strength, at which the proteins bind to the column. Then the column is eluted with a gradient of increasing ionic strength. Fractions are collected during the gradient elution. If the equipment is available, an automated FPLC system is great. However, simpler technologies can also be used.
I am under the impression that DEAE-cellulose is no longer sold. You can substitute another ion-exchange resin such as QAE-Sephadex if you are planning open-top column chromatography, or QAE-Sepharose if you are planning to use an FPLC. Ion exchange chromatography is usually started at a low ionic strength, at which the proteins bind to the column. Then the column is eluted with a gradient of increasing ionic strength. Fractions are collected during the gradient elution. If the equipment is available, an automated FPLC system is great. However, simpler technologies can also be used.
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Dr Mugdha,
I don't have FPLC. How can I proceed? Could you please suggest me.
Dr Mugdha,
I don't have FPLC. How can I proceed? Could you please suggest me.
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VOTE
DEAE Cellulose still sold through Santa Cruz
https://www.scbt.com/scbt/product/deae-cellulose-9013-34-7
DEAE Cellulose still sold through Santa Cruz
https://www.scbt.com/scbt/product/deae-cellulose-9013-34-7
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VOTE
If you have a small peristaltic pump, a magnetic stirrer, a 2-chamber gradient former, and some appropriately sized flexible tubing, you can do ion-exchange chromatography without an FPLC.
If you don't have a peristaltic pump, you can use gravity to elute the column.
If you don't have a gradient former, you can use 2 beakers connected with tubing. You can also avoid making a continuous gradient and instead use a series of small step elutions of gradually increasing salt concentration.
An automatic fraction collector is nice, but if you don't have that you can collect fractions by hand.
If you have a small peristaltic pump, a magnetic stirrer, a 2-chamber gradient former, and some appropriately sized flexible tubing, you can do ion-exchange chromatography without an FPLC.
If you don't have a peristaltic pump, you can use gravity to elute the column.
If you don't have a gradient former, you can use 2 beakers connected with tubing. You can also avoid making a continuous gradient and instead use a series of small step elutions of gradually increasing salt concentration.
An automatic fraction collector is nice, but if you don't have that you can collect fractions by hand.
More
VOTE
i would like to suggeest sphedax.
i would like to suggeest sphedax.
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Just to answer your question DEAE Cellulose is also available through the following Company:
http://srlchem.com/search/search_results/productName/deae%20cellulose
Just to answer your question DEAE Cellulose is also available through the following Company:
http://srlchem.com/search/search_results/productName/deae%20cellulose
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VOTE
If you have a small peristaltic pump, a magnetic stirrer, a 2-chamber gradient former, and some appropriately sized flexible tubing, you can do ion-exchange chromatography without an FPLC.
If you don't have a peristaltic pump, you can use gravity to elute the column.
If you don't have a gradient former, you can use 2 beakers connected with tubing. You can also avoid making a continuous gradient and instead use a series of small step elutions of gradually increasing salt concentration.
An automatic fraction collector is nice, but if you don't have that you can collect fractions by hand.
If you have a small peristaltic pump, a magnetic stirrer, a 2-chamber gradient former, and some appropriately sized flexible tubing, you can do ion-exchange chromatography without an FPLC.
If you don't have a peristaltic pump, you can use gravity to elute the column.
If you don't have a gradient former, you can use 2 beakers connected with tubing. You can also avoid making a continuous gradient and instead use a series of small step elutions of gradually increasing salt concentration.
An automatic fraction collector is nice, but if you don't have that you can collect fractions by hand.
More
VOTE