ProteoSpin™ Abundant Serum Protein Depletion Kit
For the rapid depletion of abundant proteins from serum samples
For research use only and NOT intended for in vitro diagnostics.
ProteoSpin™ Abundant Serum Protein Depletion Kit
For the rapid depletion of abundant proteins from serum samples
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Features and Benefits
- Rapid and efficient removal of abundant proteins from serum and plasma samples
- Process 10 samples in 30 minutes
- Allows for visualization of low abundance proteins
- Convenient and affordable spin column protocol
- Generic protocol allows for depletion from human and other animal samples
- Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
This kit provides a fast and simple procedure for the effective depletion of major serum proteins including albumin, α-antitrypsin, transferrin and haptoglobin from serum and plasma samples. Such abundant proteins usually obscure less abundant proteins in gel electrophresis making them difficult to visualize and recover. The kit reduces sample complexity to allow for convenient investigation of less abundant proteins. In other applications such as mass spectroscopy, fractionating abundant proteins improves the resolution of less abundant proteins. The kit is 'generic' in that it is not antibody-based and can thus be used to deplete abundant proteins from any starting material - human or various other animals - making it more versatile, flexible and affordable than other kits. Resulting proteins are ready for a range of applications including 2D gel electrophoresis, SDS-PAGE, Mass Spectrometry, Protein microarrays, DIGE (Difference Gel Electrophoresis) and more.
Details
Supporting Data
Figure 1. Depletion of Abundant Proteins. Abundant proteins were depleted from human serum using the ProteoSpinTM Abundant Serum Protein Depletion Kit. Lane 1 represents the input human serum proteins, while Lanes 3 and 4 represent the elution after the abundant proteins were depleted with the kit. Lane 2 represents the binding flowthrough, and contains the proteins that have been depleted from the serum sample.
Figure 2. Depletion of Abundant Proteins. A 2D gel was run after the abundant serum proteins were depleted from human serum using the ProteoSpinTM Abundant Serum Protein Depletion Kit. The abundant proteins including albumin, a-antitrypsin, transferin and haptoglobin have been removed from the sample, allowing the visualization and detection of the lower abundance proteins of interest.
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Kit Specifications
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| Maximum Column Capacity |
500 μg
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| Minimum Column Capacity |
200 μg
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| Minimum Elution Volume |
30 μL
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| Time to Process 10 Samples |
30 minutes
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Storage Conditions
All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 2 years after the date of shipment.
Component | Cat. 17300 (25 preps) |
|---|---|
Wash Solution G | 125 mL |
Elution Buffer C | 8 mL |
Protein Neutralizer | 4 mL |
Spin Columns | 25 |
Collection Tubes | 25 |
Elution Tubes (1.7 mL) | 25 |
Product Insert | 1 |
Documentation
FAQs
Spin Column
The protein solution may not be flowing through the column due to one or more of the following:
- Centrifugation speed was too low.
Check the centrifuge to ensure that it is capable of generating 6,700 x g. Sufficient centrifugal force is required to move the liquid phase through the resin.
- Protein solution is too viscous.
Dilute the protein solution as described in the protocol.
- Cellular debris is present in the protein solution.
Filter the sample using a 0.45 µM filter or spin down insoluble materials and transfer the liquid portion to the column. Solid, insoluble materials can cause severe clogging problems.
- Inadequate spin time.
Spin for an additional minute to ensure that the liquid is able to flow completely through the column.
Insufficient depletion of major proteins may be caused due to:
- Overloading the column with proteins.
Decrease the amount of serum or plasma that is loaded onto the column.
- Improper sample preparation.
Ensure that the serum or plasma sample is properly prepared by diluting it in the provided Wash Solution G.
The eluted protein may be degraded due to one or more of the following:
- Eluted protein solution was not neutralized.
Add 18.6 µL of Protein Neutralizer to each 200 µL of eluted protein in order to adjust the pH to neutral. Some proteins are sensitive to high pH, such as the Elution Buffer C at pH 12.5.
- Eluted protein was not neutralized quickly enough.
If eluted proteins are not used immediately, degradation will occur. We strongly suggest adding Protein Neutralizer in order to lower the pH.
- Proteases may be present.
Use protease inhibitors during all steps of sample preparation, and during storage of the serum, if desired.
- Bacterial contamination of the protein solution.
Prepare the serum samples with 0.015% sodium azide.
Low protein concentration in the elution can be caused by a low level of serum proteins present in the initial sample. Increase the amount of serum or plasma that is loaded onto the column. The input amount can be verified using a reliable colorimetric assay. We recommend starting with 600 μg to 800 μg of protein in input sample.
Citations
| Title | Chi-Yu Lu, Chia-Hsien Feng |
| Citation | Journal of Pharmaceutical and Biomedical Analysis 2023. |
| Authors | Chi-Yu Lu, Chia-Hsien Feng |
| Title | Clinical Proteomics: Liquid Chromatography-Mass Spectrometry (LC-MS) Purification Systems |
| Citation | Protein Chromatography 2023. |
| Authors | Henry, M., & Meleady, P |
| Title | Doublecortin-Like Kinase 1 Is Elevated Serologically in Pancreatic Ductal Adenocarcinoma and Widely Expressed on Circulating Tumor Cells |
| Citation | PLoS One 2023. |
| Authors | D Qu, J Johnson, P Chandrakesan, N Weygant, R May, N Aiello, A Rhim, L Zhao, W Zheng, S Lightfoot, S Pant, J Irvan, R Postier, J Hocker, JS Hanas, N Ali, SM Sureban, G An, MJ Schlosser, B Stanger, CW Houchen |
| Title | Identification of plasma Complement C3 as a potential biomarker for neuroblastoma using a quantitative proteomic approach |
| Citation | Journal of Proteomics 2023. |
| Authors | PY Kim, O Tan, S Diakiw, D Carter, EO Sekerye, VC Wasinger, T Liu, M Kavallaris, MD Norris, M Haber, L Chesler, A Dolnikov, TN Trahair, N Cheung, GM Marshall, BB Cheung |
| Title | Neonatal NET-inhibitory factor and related peptides inhibit neutrophil extracellular trap formation |
| Citation | The Journal of Clinical Investigation 2023. |
| Authors | Yost, C. C., Schwertz, H., Cody, M. J., Wallace, J. A., Campbell, R. A., Vieira-de-Abreu, A., ... & Rondina, M. T |
| Title | Proteome analysis of rat serum proteins adsorbed onto synthetic octacalcium phosphate crystals |
| Citation | Analytical Biochemistry 2023. |
| Authors | Hirofumi Kaneko, Junichi Kamiie, Hirotaka Kawakami, Takahisa Anada, Yoshitomo Honda, Naru Shiraishi, Shinji Kamakura, Tetsuya Terasaki, Hidetoshi Shimauchi, Osamu Suzuki |
| Title | Proteomics of serum proteins adsorbed onto hydroxyapatite single-crystal particles with an anisotropic structure |
| Citation | Materialia 2023. |
| Authors | Erika Onuma a, Hayato Ito b, Maki Sasaki b, Nobuyuki Kanzawa c d, Keiji Kito b, Mamoru Aizawa a d |