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SEP050 Millipore

Seppro® Supermix

LC2

  •  NACRES NA.32

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Properties

Related Categories Molecular Biology, Native Protein Sample Preparation, Protein Depletion, Proteomics
Quality Level   200
bed volume   2 mL
column D × L   6.4 mm × 63 mm
capacity   loading(Flow Through volume from IgY 14 LC5 (SEP030))
storage temp.   2-8°C

Description

General description

Immunoaffinity fractionation of highly-abundant proteins (HAP) has proven to be one of the most effective approaches for overcoming the wide dynamic range of protein concentration, as well as enabling the detection of low abundant proteins (LAP). Avian polyclonal IgY (Immunoglobulin Yolk) antibodies have unique and advantageous features that allow for highly-specific fractionation of a complex protein mixture. IgY12 column can effectively remove 12 HAP from plasma. The next level of abundant protein, moderately abundant proteins (MAP), becomes an obstacle to access LAP, where the majority of biologically interesting and clinically important biomarkers reside. Therefore, isolation of MAP is a new challenge for effectively and accurately detecting and analyzing LAP.

To tackle this challenge, we further developed our IgY columns system by establishing a SuperMix column to separate MAP from LAP. The SuperMix System was developed by immunizing chickens with a flow-through fraction of IgY12 or IgY14 column and constructing the column with affinity-purified IgY antibodies against the flow-through proteins of IgY12 or IgY14.

Application

Seppro® Supermix, LC2 (2mL column) is used to separate moderately abundant proteins (MAP) from human serum low abundant proteins (LAP) after specific human serum highly-abundant proteins (HAP) have been removed using Seppro® IgY12 or IgY14 affinity separation.

Other Notes

for removal of moderately abundant proteins from human serum or plasma

Storage and Stability

Store the column at 2-8 °C. After use, equilibrate the column with 1′ Dilution Buffer containing 0.02% sodium azide and store the column at 2-8 °C with the end-caps tightly sealed. Do Not Freeze the column.

Legal Information

Seppro is a registered trademark of Sigma-Aldrich Co. LLC

Kit component also available separately

Description

Product #

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Seppro® Dilution buffer SDS S4199
Seppro® Stripping Buffer SDS S4324
Seppro® Neutralization Buffer SDS S4449
Corning® Costar® Spin-X® centrifuge tube filters, cellulose acetate membrane, pore size 0.45 μm, non-sterile SDS CLS8163
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Frequently Asked Questions

How does the storage temperature relate to shipping conditions?
The storage conditions that a Sigma-Aldrich catalog and label recommend for products are deliberately conservative. For many products, long-term storage at low temperatures will increase the time during which they are expected to remain in specification and therefore are labeled accordingly. Where short-term storage, shipping time frame, or exposure to conditions other than those recommended for long-term storage will not affect product quality, Sigma-Aldrich will ship at ambient temperature. The products sensitive to short-term exposure to conditions other than their recommended long-term storage are shipped on wet or dry ice. Ambient temperature shipping helps to control shipping costs for our customers. At any time, our customers can request wet- or dry-ice shipment, but the special handling is at customer expense if our product history indicates that the product is stable for regular shipment. See Shipping and Storage for more information.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 or view local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
How many moderately abundant proteins (MAPs) doesProduct SEP050, Seppro® Supermix, deplete?
As of this time, the exact number of depleted MAPs by SEP050 is not precisely defined.A study by Qian et al. (Mol. Cell. Proteomics, 7(10), 1963-1973 (2008)) indicates that at least 45 MAPs were captured.
If I want to use the SEP050 SuperMix LC2 column along with one of the IgY14 columns, do I have to use the IgY14 LC5 (SEP030)?Can I use one of the others (LC2, SEP020; LC10, SEP040)?
With SEP050, it is recommended to use the SEP030 IgY14 LC5 column, because of the scale of the run.
My protein sample contains SDS.Is this compatible with Product SEP050, Seppro® Supermix?
No. We do not recommend using protein samples containg SDS with Product No. SEP050. SDS is a detergent and would be expected to affect adversely the activity of the conjugated antibody in terms of depletion.
I originally purchased my Seppro SuperMix LC2 column from Genway.Can I still contact Genway for technical support?
All inquiries on the Seppro technology are now to be referred to Sigma-Aldrich. Customers should not contact Genway for technical assistance.
My question is not addressed here, how can I contact Technical Service for assistance?
Ask a Scientist here.
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Protocols & Articles

Articles

Development of Two Multiplex Immunoassays and Preliminary Application in Determining Protein Depletion

Depletion of Highly Abundant Protein (HAP) and Moderately Abundant Protein (MAP) from plasma or serum samples can be achieved through Sigma Seppro Tandem IgY14/ SuperMix columns. A range of 77–129 MA...
Jiaxin Dong, Chris Melm, Holly Chapman, Farrah Fan, Dian Er Chen and Henry Duewel
Protein Technology and Assay, Research Biotechnology, Sigma-Aldrich Corporation
Keywords: Enzyme-linked immunosorbent assay, Filtration, High performance liquid chromatography, Immobilization, Mass spectrometry

Development of two Multiplex Immunoassays

Depletion of Highly Abundant Protein (HAP) and Moderately Abundant Protein (MAP) from plasma or serum samples can be achieved through our Seppro® Tandem IgY14/ SuperMix columns.
Jiaxin Dong, Chris Melm, Holly Chapman, Farrah Fan, Dian Er Chen, Henry Duewel
Biowire Volume 10 Article 1
Keywords: Enzyme-linked immunosorbent assay, Filtration, High performance liquid chromatography, Immobilization, Indirect ELISA, Mass spectrometry, Proteomics

Human IgY14 and SuperMix Columns

Immunoaffinity partitioning of highly-abundant proteins (HAP) has proven to be one of the most effective approaches for overcoming the wide dynamic range of plasma protein concentration, as well as e...
Keywords: Fractionation, High performance liquid chromatography, Mass spectrometry, PAGE

Protein Depletion Multiplex Immunoassays

Depletion of Highly Abundant Protein (HAP) and Moderately Abundant Protein (MAP) from plasma or serum samples can be achieved through Sigma Seppro® Tandem IgY14/ SuperMix columns. A range of 77–129 M...
Jiaxin Dong, Chris Melm, Holly Chapman, Farrah Fan, Dian Er Chen and Henry Duewel
Protein Technology and Assay, Research Biotechnology, Sigma-Aldrich Corporation
Keywords: High performance liquid chromatography, Mass spectrometry

Teaming Seppro® and iTRAQ® for Biomarker Discovery

In proteomics, protein depletion technologies, such as Seppro from Sigma-Aldrich, have proven useful in increasing sample dynamic range via removal of high-abundance proteins that are not necessarily...
George Yeh, Product Manager, Sigma® Life Science
Biofiles Vol. 8, No. 5
Keywords: Cancer, Clinical, Mass spectrometry, Proteomics

The Human IgY14/Supermix LC1 Column

The Seppro IgY14/Supermix Combo LC1 column contains both IgY14 and Supermix resins with a ratio of 2:1. The resin can be reused up to 100 times.
Keywords: Buffers, Mass spectrometry, Vitamins

Peer-Reviewed Papers
15

References

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