SEP020

Millipore

Seppro® IgY14

LC2

NACRES:
NA.32

Quality Level

column D × L

6.4 mm × 63.0 mm

capacity

2.5 mg total protein loading(35 μL of human plasma, average human protein concentration 70 mg/mL)

storage temp.

2-8°C

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General description

The targeted highly abundant proteins are simultaneously removed by the immobilized specific IgYs when crude biological samples are passed through the column. Selective immunodepletion provides an enriched pool of low abundance proteins for downstream proteomic analyses. Specific removal of these 14 highly abundant proteins depletes ~95% of the total protein mass from human serum or plasma. The low abundance proteins in the flow-through fractions can then be studied. Removal of highly abundant proteins enables improved resolution and dynamic range for one dimensional electrophoresis (1DGE), two-dimensional (2DGE) electrophoresis and liquid chromatography/ mass spectrometry (LC/MS). The collected flow-through fractions may need to be concentrated dependent upon the downstream application.
Seppro® IgY14 columns allow users to separate 14 highly abundant proteins (HAP) from plasma protein mixtures. The Avian polyclonal IgY (Immunoglobulin Yolk) antibodies, have a distinct, highly specific partitioning advantage over competing products, allowing the researcher to look at a much cleaner human protein mixture. Seppro® columns make further proteomics analysis into the plasma proteome much easier and provide cleaner results.

Proteins Depleted:

Albumin
IgG
α1-Antitrypsin
IgA
IgM
Transferrin
Haptoglobin
α2-Macroglobulin
Fibrinogen
Complement C3
α1-Acid Glycoprotein (Orosomucoid)
HDL (Apolipoproteins A-I and A-II)
LDL (mainly Apolipoprotein B)

Application

Seppro® IgY14, LC2 (2 mL bed volume columns) is designed for use as a plasma/serum protein removal tool base upon immune affinity removal of 14 highly abundant proteins (HAP) from human plasma protein mixtures via liquid chromatography. Seppro® IgY14 is used to support the analysis of low abundance human serum/plasma proteins.

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.

Other Notes

If the protein concentration of the sample is unknown and the total serum protein level is potentially elevated, a reduction of the serum load to 24.0 ml is recommended for initial study to avoid potential abundant protein bleed through.

Legal Information

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

Kit Components Also Available Separately

Product No.
Description
SDS

  • S4199Seppro® Dilution buffer

  • S4324Seppro® Stripping Buffer

  • S4449Seppro® Neutralization Buffer

  • CLS8163Corning® Costar® Spin-X® centrifuge tube filters, cellulose acetate membrane, pore size 0.45 μm, non-sterile

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Precautionary Statements

RIDADR

NONH for all modes of transport

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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. 

  6. How would I decide between Products SEP020, Seppro® IgY, and PROT20LC, ProteoPrep® 20 Plasma Immunodepletion LC Columns?

    SEP020 is designed to deplete the top 14 abundant human proteins, while PROT20LC is designed to deplete the top 20 abundant human proteins. Also, all of the antibodies in SEP020 are IgY antibodies, while the PROT20LC antibodies and antibody ligands are primarily IgG-based.

  7. My protein sample contains SDS. Is this compatible with Product SEP020, Seppro® IgY14?

    No. We do not recommend using protein samples containing SDS with Product No. SEP020. SDS is a detergent and would be expected to affect adversely the activity of the conjugated antibody in terms of depletion.

  8. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

  9. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Effect of plasma protein depletion on BNP-32 recovery.
Adam M Hawkridge et al.
Clinical chemistry, 54(5), 933-934 (2008-04-30)
Does plasma protein depletion increase lung liquid conductance?
P Dodek
American journal of respiratory and critical care medicine, 154(4 Pt 1), 1213-1215 (1996-10-01)
Renato Millioni et al.
PloS one, 6(5), e19603-e19603 (2011-05-17)
To date, the complexity of the plasma proteome exceeds the analytical capacity of conventional approaches to isolate lower abundance proteins that may prove to be informative biomarkers. Only complex multistep separation strategies have been able to detect a substantial number...

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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