Three Research Articles Cite Cleanascite™ in Bile Proteomics

The first citation is:

Annarita Farina, Jean-Marc Dumonceau, Paola Antinori, Isabelle Annessi-Ramseyer, Jean-Louis Frossard, Denis F. Hochstrasser, Myriam Delhaye, Pierre Lescuyer. Bile carcinoembryonic cell adhesion molecule 6 (CEAM6) as a biomarker of malignant biliary stenoses. Biochimica et Biophysica Acta 1844 (2014) 1018–1025

In brief, the article’s authors aimed at identifying new cancer biomarkers by comparing proteomic analysis of bile collected from patients with a malignant or benign biliary stenosis. The article states “For sample preparation, bile samples were centrifuged at 16,000g for 10 minutes. Each supernatant was delipidated with Cleanascite…”

The second citation is:

Natalija Lukic, Rémy Visentin, Myriam Delhaye, Jean-Louis Frossard, Pierre Lescuyer,
Jean-Marc Dumonceau, Annarita Farina. An integrated approach for comparative proteomic analysis of human bile reveals overexpressed cancer-associated proteins in malignant biliary stenosis. Biochimica et Biophysica Acta 1844 (2014) 1026–1033

In brief, the article’s authors acknowledged that the complexity of bile and the presence of molecules interfering with protein analysis represent an obstacle for quantitative proteomic studies in bile samples. In this study, differential centrifugation was used as a preliminary step in a quantitative proteomic workflow involving iTRAQ labeling.

Sample preparation for proteomic analysis was as follows:
“Bile samples were centrifuged at 16,000g for 20 min at 4C. The obtained supernatants were diluted (1:5) with Dulbecco’s Phosphate Buffered Saline (D-PBS) to reduce viscosity and subjected to the following consecutive steps of differential centrifugation: i) 70.000 g for 60 min; ii) 100,000 g for 60 min; and iii) 200,000 g for 120 min at 4C. The pellet fractions obtained after each centrifugation step were washed with an equal volume of D-PBW and centrifuged again, under the same conditions to remove residual supernatant. Equal aliquots of each supernatant fraction, corresponding to the same initial volume (45ul) of crude bile, were delipidated with Cleanascite.” A total of 1267 proteins were identified, including 322 newly described bile proteins that the authors conclude were mainly attributable to high density cellular fractions.

The third citation is:

Elisa Danese, PhD, Orazio Ruzzenente, BS, Andrea Ruzzenente, MD, PhD, Calogero Iacono, MD, Francesca Bertuzzo, MD, Matteo Gelati, MT, Simone Conci, MD, Sharon Bendinelli, MT, Giada Bonizzato, MT, Alfredo Guglielmi, MD, Gian Luca Salvagno, MD Giuseppe Lippi, MD, and Gian Cesare Guidi, MD. Assessment of bile and serum mucin5AC in cholangiocarcinoma: Diagnostic performance and biologic significance.

The author’s aim was to improve the biologic relevance of the mucin MUC5AC in malignant and benign biliary disorders by comparing its diagnostic performance in both bile and serum samples of patients with cholangiocarcinoma (CCA) or benign biliary disorders.
Delipidation of bile was performed as follows: “After centrifugation, the supernatant of each sample was mixed with 250 ul ofCleanascite (v/v ratio Cleanascite per sample = 1:4) and kept under mild agitation at 4C for 1 hour…”

“I am excited by the number of recent research articles on bile proteomics; 7 in the last 2 years alone. Clearly the excellent selectivity profile that Cleanascite™ provides is making its use routine whenever bile characterization requires complex analysis.” states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.

Cleanascite™ selectively removes lipids, cell debris, lipoproteins, impurities from cohn paste, transgenic milk, egg yolk and biological samples for pretreatment and clarification of samples. The reagent is a solid-phase, non-ionic adsorbent supplied as a suspension in saline, ready for use. Simply add, centrifuge and/or filter. The clarified supernatant is ready for subsequent downstream processing or analysis.

• A high binding capacity for lipids with minimal cross-reactivity with proteins
• Effectively replaces chlorinated/fluorinated hydrocarbons (eg. freon) with no environmental hazards.
• Helps purify antibodies, recombinant proteins, nucleic acids, proteoglycans
• Ideal for clarifying ascites, serum, cell & tissue culture, bile and organ homogenates
• Clarifies saliva and fecal components
• Excellent selectivity profile, especially suitable for LC-MS proteomics
• Extends the life of membrane and chromatographic columns

For more information visit: Cleanascite™ Lipid Removal Reagent and Clarification, at
http://www.biotechsupportgroup.com/cleanascite-lipid-removal-and-clarification

About Biotech Support Group LLC
Biotech Support Group LLC is a leading provider of proteomic, metabolomic and genomic sample preparation and enrichment products and services. It’s principal products include: AlbuVoid™ & AlbuSorb™ for albumin depletion, Cleanascite™ for lipid adsorption and clarification, HemogloBind™ & HemoVoid™ for hemoglobin removal, NuGel™ for functional & chemical proteomics, and ProCipitate™ for nucleic acid isolation. For more information, go to http://www.biotechsupportgroup.com

Contact:
Matthew Kuruc & Dr.Swapan Roy
mkuruc@biotechsupportgroup.com
Biotech Support Group LLC
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