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  5. Thermo Scientific™ High Select™ Depletion Spin Columns

Thermo Scientific™ High Select™ Depletion Spin Columns

Catalog No. PIA36372
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Quantity:
10 Columns
24 Columns
6 Columns
50 mL
Product Type:
Midi Spin Column
Mini Spin Column
Resin
Purification Target:
HSA/Immunoglobulin
Top14 Abundant Protein
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Catalog No. Quantity Product Type Purification Target
PIA36372 50 mL Resin Top14 Abundant Protein
PIA36365 6 Columns Mini Spin Column HSA/Immunoglobulin
PIA36366 24 Columns Mini Spin Column HSA/Immunoglobulin
PIA36367 10 Columns Midi Spin Column HSA/Immunoglobulin
PIA36368 50 mL Resin HSA/Immunoglobulin
PIA36371 10 Columns Midi Spin Column Top14 Abundant Protein
PIA36369 6 Columns Mini Spin Column Top14 Abundant Protein
PIA36370 24 Columns Mini Spin Column Top14 Abundant Protein
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Catalog No. PIA36372 Supplier Thermo Scientific™ Supplier No. A36372
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Description

Decrease albumin, immunoglobulins, and other high-abundance proteins from human serum or plasma in preparation for mass spectrometry analysis, or 1D and 2D gel electrophoresis using High Select depletion spin columns or resin.

Analysis of human fluids is often complicated by the presence of high concentrations of serum proteins, including albumin and IgG that can make up more than 70% of total serum protein. Removal of albumin, immunoglobulins, and other abundant proteins is essential for the study of low-abundance proteins. Thermo Scientific High Select HSA/Immunoglobulin and Top 14 Abundant Protein depletion spin columns and resins enable depletion of the most abundant proteins in human serum and plasma for improved detection of low-abundance proteins using mass spectrometry analysis or 1D or 2D gel electrophoresis.

• Flexible—resin provided in mini and midi column formats as well as bulk for custom filling options
• Optimized—removes >95% of IgG and >95% of albumin (HSA/Immunoglobulin); removes >95% of IgG and >95% of albumin, plus 12 other abundant proteins (Top 14)
• Fast—process samples in ∼10 minutes
• Economical—cost-effective spin columns are priced for single use
• Reduced carry-over—one-time use prevents protein carryover and experimental variability

High Select HSA/Immunoglobulin Depletion Resin uses highly specific, immobilized anti-HSA and anti-immuoglobulin antibodies (IgG, IgA, IgM, IgD, and IgE) to remove human serum albumin (HSA) and all major subclasses of immunoglobulins from serum and plasma. High Select Top14 Abundant Protein Depletion Resin uses highly specific immobilized antibodies to remove human serum albumin (HSA), albumin, IgG, IgA, IgM, IgD, IgE, kappa and lambda light chains, alpha-1-acidglycoprotein, alpha-1-antitrypsin, alpha-2-macroglobulin, apolipoprotein A1, fibrinogen, haptoglobin, and transferrin from serum, plasma, or spinal fluids. The resins are provided in an economical and convenient spin column format specifically designed for one-step processing and single use or bulk format for custom filling.

Specifications

For Use With (Equipment) Mass Spectormeter
Product Type Resin
Product Line High Select
Content And Storage Store at 2–8°C.
Label or Dye Unlabeled
Shipping Condition Wet Ice
Starting Material Plasma, Serum
Final Product Type Protein
Workflow Step Protein Depletion
Quantity 50 mL
Purification Target Top14 Abundant Protein
Form Liquid
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Frequently Asked Questions (FAQs)

Is there a 96-well format available for the Top14 Abundant Protein Depletion Mini Spin Columns (Cat. No. A36369)?

No. The Top14 Abundant Protein Depletion Mini Spin Columns (Cat. No. A36369) do not come in a 96-well format. However, we do have a Top14 Abundant Protein Depletion Resin (Cat. No. A36372) that can be purchased as a stand-alone product.

After depletion of abundant proteins from my sample, I still see albumin and other abundant proteins after LC-MS analysis. Did the depletion work?

Depletion does not remove all abundant proteins from the sample. Even with 99% depletion of a specific plasma protein, highly abundant proteins will still be the most abundant proteins in the sample detected by MS analysis. However, depletion significantly decreases the most abundant plasma proteins enabling identification of lower abundant proteins.

How do I proceed to downstream mass spec sample preparation after abundant plasma protein depletion?

After the depletion, samples are significantly diluted in buffer. Depleted samples should be concentrated by speed vac drying, solvent precipitation, or diafiltration. Samples dried using a speedvac are directly compatible with the EasyPep kit chemistry after resuspension using the Lysis Solution.
What is the maximum loading capacity for the High Select Depletion columns?

Mini columns can deplete up to 10 µL (600 µg) of plasma or serum; midi columns can deplete up to 100 µL (6000 µg) of plasma or serum.
What is the percentage of albumin that is removed using abundant protein depletion columns?

More than 99% of albumin can be depleted when columns use the recommended sample to resin ratio.
How do I assess efficiency and reproducibility of abundant protein depletion in plasma samples?

ELISA and mass spectrometry analysis are often used for evaluation of depletion performance. Protein yields after depletion can be measured using the BCA assay.

What formats of the High Select Top 14 Abundant Protein Depletion Resin do you offer?

The High Select Top 14 Abundant Protein Depletion Resin is offered in 3 formats:

a) Resin alone: High Select Top 14 Abundant Protein Depletion Resin (Cat. No. A36372)
b) High Select Top 14 Abundant Protein Depletion Mini Spin Columns (Cat. Nos. A36369, A36370) that can process a maximum of 10 µL of human serum or plasma
c) High Select Top 14 Abundant Protein Depletion Midi Spin Columns (Cat. No. A36371) that can process a maximum of 100 µL of human serum or plasma. For serum samples containing abnormally high amounts of albumin or IgG, the sample load may need to be reduced.
Which proteins are removed by High Select Top 14 Abundant Protein Depletion Resin?

The High Select Top14 Abundant Protein Depletion Resin uses immobilized antibodies to remove human serum albumin (HSA), albumin, IgG, IgA, IgM, IgD, IgE, kappa and lambda light chains, alpha-1-acidglycoprotein, alpha-1-antitrypsin, alpha-2-macroglobulin, apolipoprotein A1, fibrinogen, haptoglobin, and transferrin from serum, plasma, or spinal fluids.