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Pierce™ Phosphoprotein Enrichment Chromatography Cartridges, 5mL - FAQs

查看更多产品信息 Pierce™ Phosphoprotein Enrichment Chromatography Cartridges, 5mL - FAQs (87744)

9 个常见问题解答

Can I recharge the column in the Thermo Scientific Phosphoprotein Enrichement kit?

No. The metal and procedure used to charge the columns are proprietary.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Can I reuse or recharge the columns when using the Thermo Scientific Phosphoprotein Enrichment kit?

No. For best results, use the column only one time.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Are samples containing EDTA or reducing agents compatible with the Thermo Scientific Phosphoprotein Enrichment kit?

No. Chelating agents such as EDTA and EGTA will cause metal to leach from the column and hence, reduce binding. Reducing agents such as DTT, 2-mercaptoethanol and TCEP also reduce binding to the column. Remove incompatible substances from the sample by desalting before applying to the column.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Do I need phosphatase inhibitors when using the Thermo Scientific Phosphoprotein Enrichment kit?

Phosphatase and protease inhibitors are not required. If desired, add inhibitors to the Lysis/Binding/Wash Buffer as indicated in the product instructions.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What other type of lysis buffer can I use with the Thermo Scientific Phosphoprotein Enrichment kit?

You can use RIPA buffer to lyse cells; however, dilute the lysate with the Lysis/Binding/Wash Buffer included in the kit for compatibility with the column.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What type of yield should I expect with the Thermo Scientific Phosphoprotein Enrichment kit?

Typical yields are 10-25% of the total protein, depending on cell stimuli, cell type and binding characteristics of the specific phosphoproteins.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

How many samples can I process and what is included in the Thermo Scientific Phosphoprotein Enrichment kit?

Each of the 10 columns in the kit can process a single sample containing up to 4 mg of total protein. The kit includes Lysis/Binding/Wash Buffer, CHAPS, Elution Buffer and 10 Thermo Scientific Protein Concentrators (7 mL/9K) for concentrating the enriched samples.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Will the columns bind phosphoproteins modified on tyrosine residues?

Yes. The Phosphoprotein Enrichment Columns bind phosphotyrosine, phosphoserine and phosphothreonine residues.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

How does the Thermo Scientific Phosphoprotein Enrichment kit work?

The columns in the Thermo Scientific Phosphoprotein Enrichment Kit contain a proprietary metal that interacts with negative charges from phosphate groups. The optimized buffer conditions enable specific capture of phosphoproteins from complex biological samples.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.