Capturem high-capacity membranes

Capturem His-tagged 24-Well Plates are 24-well plates that provide rapid, high-quality, small-scale purification of his-tagged proteins. These plates combine Ni-NTA chemistry with novel Capturem technology to provide high-capacity, membrane-based affinity purification. Typical yields for his-tagged proteins are up to 800 µg per well with a 4.5-ml sample capacity per well.

Capturem His-Tagged Purification 96 is a single-use, disposable 96-well plate for simple, rapid purification of his-tagged proteins from up to 1 ml of clarified lysate per well. This plate is suitable for use under native or denaturing conditions, in the presence of additives such as DTT (up to 10 mM), βME (up to 30 mM), TCEP (up to 5 mM), EDTA (up to 10 mM), or glycerol. Capturem His-Tagged Purification 96 is compatible with automated liquid handlers equipped with vacuum filtration or centrifuges equipped with appropriate plate holders.

The Capturem His-Tagged Purification Large Volume lets you quickly purify up to 25 mg of protein (depending on protein size and expression level) from a large volume of lysate, (up to 500 ml at a time) resulting in purified protein concentrated at up to 2 mg/ml. Like our other his-tagged Capturem products, the Large Volume unit works for bacterial and mammalian expression systems with a wide range of additives.

Capturem His-Tagged Purification Maxiprep Columns are single-use disposable membrane spin columns for simple, rapid purification of his-tagged proteins in up to 25 ml of clarified lysate. The columns are suitable for use under native or denaturing conditions, and in the presence of additives such as DTT (up to 10 mM), βME (up to 30 mM), TCEP (up to 5 mM), EDTA (up to 10 mM), or glycerol.

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High yield and activity of his-tagged Metridia luciferase purified using the Capturem His-Tagged Purification Maxiprep Kit

High yield and activity of his-tagged Metridia luciferase purified using the Capturem His-Tagged Purification Maxiprep Kit. Panel A. Confirmation of Metridia luciferase expression in 293T cells. The pEF1a-Metluc-6xHis-IRES2-DsRed-Express2 expression vector, which coexpresses 6xhis-tagged Metridia luciferase with a red fluorescent reporter, was constructed using In-Fusion Cloning and the pEF1alpha-IRES-DsRed-Express2 Vector, and transfected into 293T cells using Xfect Transfection Reagent. The DsRed-Express2 reporter was then used to monitor transfection efficiency. Panel B. Yield and activity of Metridia luciferase, pre- and post-purification. Luciferase activity was measured at various steps in the purification of culture supernatant prepared from 293T cells expressing his-tagged Metridia luciferase.

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Capturem his-tagged maxiprep

Capturem his-tagged maxiprep. Capturem his-tagged maxiprep spin columns have a capacity of 2.5 mg his-tagged protein from a starting culture of 25 ml.

The Capturem His-Tagged Purification Maxiprep Kit provides single-use disposable membrane spin columns for simple, rapid purification of his-tagged proteins in up to 25 ml of clarified lysate. The columns are suitable for use under native or denaturing conditions, and in the presence of additives such as DTT (up to 10 mM), βME (up to 30 mM), TCEP (up to 5 mM), EDTA (up to 10 mM), or glycerol.

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High yield and activity of his-tagged Metridia luciferase purified using the Capturem His-Tagged Purification Maxiprep Kit

High yield and activity of his-tagged Metridia luciferase purified using the Capturem His-Tagged Purification Maxiprep Kit. Panel A. Confirmation of Metridia luciferase expression in 293T cells. The pEF1a-Metluc-6xHis-IRES2-DsRed-Express2 expression vector, which coexpresses 6xhis-tagged Metridia luciferase with a red fluorescent reporter, was constructed using In-Fusion Cloning and the pEF1alpha-IRES-DsRed-Express2 Vector, and transfected into 293T cells using Xfect Transfection Reagent. The DsRed-Express2 reporter was then used to monitor transfection efficiency. Panel B. Yield and activity of Metridia luciferase, pre- and post-purification. Luciferase activity was measured at various steps in the purification of culture supernatant prepared from 293T cells expressing his-tagged Metridia luciferase.

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Capturem his-tagged maxiprep

Capturem his-tagged maxiprep. Capturem his-tagged maxiprep spin columns have a capacity of 2.5 mg his-tagged protein from a starting culture of 25 ml.

The Capturem His-Tagged Purification Miniprep Kit provides single-use disposable membrane spin columns for simple, rapid purification of his-tagged proteins in up to 800 μl of clarified lysate. The columns are suitable for use under native or denaturing conditions, in the presence of additives such as DTT (up to 10 mM), βME (up to 30 mM), TCEP (up to 5 mM), EDTA (up to 10 mM), or glycerol.

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Protein purification miniprep workflow

Protein purification miniprep workflow. Each mini spin column can be loaded with up to 800 μl of lysate (yielded from 2–5 ml of culture). His-tagged protein is first bound to the membrane, followed by washing with 300 μl wash buffer, and elution with 300 μl elution buffer. Over 90% of the bound protein can be eluted with as little as 100 μl elution buffer. Each step is followed by spinning the tube for 1 min at 11,000 x g. The working bed volume of the membrane is <2 μl. This entire purification is complete in <5 min.

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Purification of GFPuv in the presence of a wide range of additives, at varying concentrations

Purification of GFPuv in the presence of a wide range of additives, at varying concentrations. Additives were included in the sample, equilibration, wash, and elution buffers for each experiment. Samples were eluted twice with 300 µl elution buffer each time.

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Purification of GFPuv under native and denaturing conditions

Purification of GFPuv under native and denaturing conditions. Spin columns were loaded with 800 µl of cell lysate, and all steps were performed with appropriate buffers for native and denaturing conditions. 8 M urea or 6 M guanidine were included in appropriate samples.

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Purification of 6xhis-tagged proteins expressed in mammalian cells

Purification of 6xhis-tagged proteins expressed in mammalian cells. 293T cells were transfected with plasmids encoding 6xhis-tagged MAPK1 or ADRB2. Cells were lysed, and the clarified lysate was loaded onto an equilibrated column. The column was washed with 400 μl wash buffer, followed by elution with 300 μl elution buffer. The various fractions were then resolved on a 4–20% polyacrylamide gel, and blotted onto a nitrocellulose membrane. Panel A, left. Ponceau S staining of the MAPK1 membrane shows all the proteins present in the different fractions. Panel A, right. Immunoblot using an antibody specific to MAPK1 shows a band corresponding to MAPK1. Panel B, left. Ponceau S staining of the ADRB2 membrane shows all the proteins present in the different fractions. Panel B, right. Immunoblot using an antibody specific to ADRB2 shows the enrichment of ADRB2 in the eluate.

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Miniprep spin columns have a capacity of >100 μg his-tagged protein from a starting culture of 5 ml

Miniprep spin columns have a capacity of >100 μg his-tagged protein from a starting culture of 5 ml.

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