SEP050

Millipore

Seppro® Supermix

LC2

NACRES:
NA.32

Quality Level

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

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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 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 many moderately abundant proteins (MAPs) does Product 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.

  7. 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.  

  8. 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.

  9. 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.

  10. 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)
Majlinda Kullolli et al.
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 939, 10-16 (2013-10-05)
Human plasma is a commonly used diagnostic fluid in clinical chemistry. In-depth plasma proteomic analysis is performed to search for disease biomarkers, however the large dynamic range of protein abundance in plasma presents a substantial analytical challenge. Removal of abundant...

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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