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Albumin and IgG Depletion From Serum/Plasma for Proteomics
- IgG removal >90% (70-80% of total Immunoglobulins removed)
- Albumin removal >95%
- Seamless and simple < 1 hour protocol
- Low abundance enrichment equivalent to immuno-affinity
- Disposable, cost-effective, no column regeneration or cross-contamination
- Works for most species tested including human, sheep, rat, mouse, bovine
- On-bead protocols improve workflow and efficiency, especially suited to targeted proteomics
- Suitable for LC-MS, 1 and 2D Gels, ELISAs, Enzyme and other Functional Assays.
The classical plasma proteins generally fall into functional categories, forming the vast majority of the mid-to-high abundance proteome. In serum, these sub-proteomes by mass content are: Albumin 50-60%; Immunoglobulins 10-20%; Transport Proteins (Transferrin, Apo) 5-10%; Complement related Proteins 3-5%; Protease Inhibitors 2-5%; and all others 2-5%. While these sub-proteomes are required for normal body homeostasis, they nevertheless can become dysfunctional during acute-phase and chronic stimuli.
So, depending on the needs of the investigation, it can be valuable to consider that one or more of these categorical sub-proteomes is simply background noise whereby depletion is beneficial. While in other cases, these same categorical sub-proteomes might provide new data and information and consequently, should not be depleted. Different AlbuVoid™, AlbuVoid™ PLUS and AlbuSorb™ PLUS workflows support different proteomic biases as outlined in the following Table.
Products and digest conditions produce different sub-proteome windows of observation. So, depending on the needs of the investigation, it can be valuable to consider that one or more of these categorical sub-proteomes is simply background noise whereby depletion is beneficial. While in other cases, these same categorical sub-proteomes might provide new data and information and consequently, should not be depleted. Categorically the acute-phase sub-proteomes differentiated in disease may vary greatly from those associated with chronic sub-proteomes. So there is great benefit in having options to enrich or deplete one or more of these sub-proteomes.
BSG's Albumin and IgG Removal Kits offer many such options:
- The 'PLUS' products substantially deplete Immunoglobulins through separations at the protein level.
- The variable regions of Immunoglobulins are extremely heterogeneous, generating a background noise across the full LC gradient. On-bead digestion (BASP™) with AlbuVoid™ substantially reduces the influence of such Ig peptide features. So in addition to workflow simplicity, BASP™ can be advantageous utilized in targeted proteomic workflows whenever the target proteins do not require strong denaturing conditions.
- With the exception of Immunoglobulins whereby FASP generates many more spectral features, both strong denaturing conditions (FASP) and on-bead digest (BASP™) conditions produce similar protein profiles.
- Both Apolipoproteins and heavily glycosylated proteins (i.e., α1-Acid Glycoprotein) bind poorly to AlbuVoid™. For quantitative studies within these classes of proteins, AlbuSorb™ PLUS is recommended.
- The Complement sub-proteome is especially enriched by AlbuVoid™ PLUS. The digest conditions may bias towards one or more functional sub-populations, likely due to conformational transitions and protein-protein interactions (i.e., Factor Bb, Properdin) that occur upon activation. This needs further investigation.
- The low abundance sub-proteome is enriched 5+ fold with AlbuVoid™ and 4+ fold with AlbuSorb™ PLUS.
Product
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Size
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# Serum Preps
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Item No.
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AlbuVoid™ PLUS Kit
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5 preps
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5, 25µl Serum samples
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NP-AVK-05
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AlbuVoid™ PLUS Kit
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10 preps
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10, 25µl Serum samples
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NP-AVK-10
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Click Here For The AlbuVoid™ Plus Product Sheet.
Items
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Item No.
NP-AVK-05
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Item No.
NP-AVK-10
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Reagents
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Kit#1 - IgG Depletion Kit
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NuGel™ Protein A Beads (NP)
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300mg
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600mg
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Supplied
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Buffer 1
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5 ml
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10 ml
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Supplied
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Spin-X Centrifuge tube Filters
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5
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10
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Supplied
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Items
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Item No.
NP-AVK-05
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Item No.
NP-AVK-10
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Reagents
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Kit#2 - Albumin Depletion Kit
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AlbuVoidTM Beads (AVK)
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125mg
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250mg
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Supplied
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Binding Buffer AVBB pH 6.0
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3 ml
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6 ml
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Supplied
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Wash Buffer AVWB pH 7.0
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12 ml
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24 ml
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Supplied
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Elution Buffer AVEB pH 10
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2 ml
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4 ml
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Supplied
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Spin-X Centrifuge tube Filters
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5
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10
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Supplied
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DTT, Iodoacetamide, Trypsin and Formic Acid
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Not Supplied
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Typical Performance
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AlbuVoid™ PLUS
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Serum Sample Volume
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25 –
50 µl
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Albumin Removal
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>95%
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IgG Removal (most species)
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>90%
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Total Immunoglobulin Removal (most species)
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70-80%
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Recoverable Protein Mass
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150 - 300 µg
(Albumin + IgG depleted)
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LC-MS/MS unique proteins (>2 Sp. Ct)
(single 2 hr gradient)
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300 - 500
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LC-MS/MS unique peptide spectral counts
(single 2 hr gradient)
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20,000 – 30,000
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For targeted proteomics, please contact technical services, as we have a knowledgebase of over 1000 serum proteins to help select the best method(s) for particular protein(s).
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References
AlbuVoid™ PLUS & AlbuSorb™ PLUS - Evaluating Different Windows of Observation Solves The Many Challenges of Serum Proteomics
Zheng H, Soherwardy A, Roy S, Kuruc M, Avadhani S. AlbuVoid™ Enrichment & Antibody Depletion – Tackling the Challenges of Serum Proteomics Part II. Poster reprint first presented at 26th International Molecular Med TRI-CON, March 10-15, 2019, San Francisco, CA USA
Vialaret, Jerome & Kadi, Sarah & Tiers, Laurent & O Flynn, Robin & Lehmann, Sylvain & Hirtz, Christophe. (2018). Albumin depletion of human serum to improve quantitative clinical proteomics. Current Topics in Peptide & Protein Research 19. 53-62.
Poillet-Perez, Laura, et al. "Autophagy maintains tumour growth through circulating arginine." Nature (2018): 1.
Roy, Swapan, and Matthew Kuruc. "Methods to Monitor the Functional Subproteomes of SERPIN Protease Inhibitors." Functional Proteomics. Humana Press, New York, NY, 2019. 41-54.
Mihara, Keisuke, et al. "Identification of Specific Protein Markers of Rheumatoid Arthritis in Synovial Fluid and Serum." Journal of Hard Tissue Biology 27.1 (2018): 55-58.
David L. Wang, Chuanguang Xiao, Guofeng Fu, Xing Wang and Liang Li. Identification of potential serum biomarkers for breast cancer using a functional proteomics technology. Biomarker Research (2017) 5:11. DOI 10.1186/s40364-017-0092-9
Zheng, H., et al. "AlbuVoid™ coupled to on-bead digestion-tackling the challenges of serum proteomics." J Proteom Bioinformatics 8.9 (2015): 225.
Zheng, H., et al. Poster reprint first presented at 14th Human Proteome Organization World Congress (HUPO 2015), held on September 27 – 30, 2015 entitled “New Proteomic Workflows Combine Albumin Depletion and On Bead Digestion, for Quantitative Cancer Serum”.
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