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  5. Thermo Scientific™ High Select™ Depletion Spin Columns

Thermo Scientific™ High Select™ Depletion Spin Columns

Product Code. 15842684 Shop All Thermo Scientific Products
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Quantity:
10 Columns
24 Columns
50 mL
6 Columns
Product Type:
Midi Spin Column
Mini Spin Column
Resin
Purification Target:
HSA/Immunoglobulin
Top14 Abundant Protein
Unit Size:
50mL
Pack of 10
Pack of 24
Pack of 6
8 product options available for selection
Product selection table with 8 available options. Use arrow keys to navigate and Enter or Space to select.
Product Code. Quantity Product Type Purification Target unitSize
15842684 6 Columns Mini Spin Column Top14 Abundant Protein Pack of 6
15802684 6 Columns Mini Spin Column HSA/Immunoglobulin Pack of 6
15812684 24 Columns Mini Spin Column HSA/Immunoglobulin Pack of 24
15822684 10 Columns Midi Spin Column HSA/Immunoglobulin Pack of 10
15832684 50 mL Resin HSA/Immunoglobulin 50mL
15862684 10 Columns Midi Spin Column Top14 Abundant Protein Pack of 10
15870863 50 mL Resin Top14 Abundant Protein 50mL
15852684 24 Columns Mini Spin Column Top14 Abundant Protein Pack of 24
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Product Code. 15842684 Supplier Thermo Scientific™ Supplier No. A36369

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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.

TRUSTED_SUSTAINABILITY

Specifications

For Use With (Equipment) Mass Spectrometer
Product Type Mini Spin Column
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
Format Mini Spin Column
Workflow Step Protein Depletion
Quantity 6 Columns
Purification Target Top14 Abundant Protein
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Frequently Asked Questions (FAQs)

Can I use the High Select Top14 Abundant Protein Depletion Midi Spin Columns to deplete proteins from rodent samples?

The High Select Top14 Abundant Protein Depletion Midi Spin Columns have only been validated for human samples. We do not recommend using them for protein depletion in rodent samples. For protein depletion in rodent samples, we recommend the following multi-species depletion resins:

  • CaptureSelect MultiSpecies Albumin Depletion Product (Cat. No. 191085305)
  • Pierce Albumin Serum Depletion Kits (Cat. No. 89875)
Are your High Select Top14 Abundant Protein Depletion Mini Spin Columns (Cat. No. A36369) compatible with cell culture media?

Our High Select Top14 Abundant Protein Depletion Mini Spin Columns have been tested and validated for protein depletion only on human plasma and serum, with each column optimized to bind up to 10 µL of sample.
We have not tested our product on cell culture media or with serum from different species, therefore we do not recommend using this product with cell culture media.
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.

I have a lot of plasma samples to prepare for mass spectrometry analysis. Is there a way to perform abundant protein depletion in a 96-well plate?

The bulk Top14 depletion resin is compatible with a 96-well filter plate such as the Agilent filter microplate (Cat. No. 200957-100). Add 600 µL of resin slurry (50%) to each of the wells in the 96-well filter plate, add 10-20 µL of sample, and incubate with gentle shaking. Depleted samples can be collected in a new 96-well polypropylene plate by centrifugation for 2 mins at 1,000 x g or by using a vacuum manifold.
Do you offer a mass spec sample prep method for low abundant proteins in samples?

High Select Top14 Abundant Protein Depletion Mini Spin columns (Cat. Nos. A36369, A36370) can be used to deplete top 14 abundant plasma/serum proteins. The depletion of highly abundant proteins from the sample enables the detection and identification of low abundant proteins. The depleted samples can be processed with EasyPep MS sample prep kit to generate digested plasma/serum samples. The clean digested samples can be processed by nanoLC-MS/MS analysis for discovery experiments or nanoLC-PRM/MS analysis for targeted experiments.

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.
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