Biotech Support Group Release: Cleanascite™ Lipid Removal Reagent and Clarification
? Effectively replaces chlorinated/fluorinated hydrocarbons (eg. freon) and it is environmentally friendly.
? 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
? Very low protein binding
? Does not bind to DNA, RNA, enzymes and proteins
? Leaves glycoproteins, antibodies, nucleic acids, hemoglobin, proteoglycans, nucleic acids, serum components(such as hormones, nutrients, globulins, clotting factors, transport proteins) alone
? Extends the life of membrane and chromatographic columns.
? Enrichment of delipidated tissue samples
? Ideal for delipidation treatments for downstream processing of large-scale therapeutic proteins, enzymes and monoclonal antibodies.
Cleanascite™ selectively removes lipids, cell debris, lipoproteins, floating fats, impurities from Cohn paste, transgenic milk, egg yolk and biological samples for pretreatment of samples prior to purification. 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.
Clarifies
? Ascites5
? Serum/Plasma2
? Bile7
? Cohn Paste
? Cell Lysates3
? Tissue Culture6
? Organ Homogenates
? Saliva/Sputum4
? Egg Yolk
? Transgenic Milk
in the purification and analysis of antibodies, proteins, nucleic acids, proteoglycans, and other macromolecules
Protocol
Supplied as an aqueous suspension of non-ionic adsorbent in saline, pH 8.0. When not in use, keep sealed. For best results store at 4?C. Do not freeze. CleanasciteTM retains full activity when stored as directed for at least 6 months.
Actual lipid concentration in biological samples can vary greatly, so the ratios shown are only intended to provide general guidance in use.
1. Resuspend CleanasciteTM by gently shaking. Excessive shaking or mixing will cause foaming. It should be completely resuspended prior to use.
2. Add 1 ml
of CleanasciteTM to 4 ml of the sample. (1 : 4 volume ratio). Mix the sample by gently shaking periodically for 10 minutes. In some cases the agglomeration of fine lipids is improved by incubation at 4?C for a minimum of one hour.
3. Centrifuge sample at 16,000 G’s for 1 minute - or - 1,000 G’s for 15 minutes.
4. Decant supernatant containing macromolecules of interest and continue with purification.
Optimization. Different sample volumes are easily scaled. Volume ratio can be adjusted up or down as required to remove the amount of impurities present. In some cases the agglomeration of fine lipids is improved by incubation at 4?C for a minimum of one hour.
References
Bile
Hauser-Davis RA, Lima AA, Ziolli RL, Campos RC.First-time report of metalloproteinases in fish bile and their potential as bioindicators regarding environmental contamination. Aquatic Toxicology.2012;110-111:99-106
Farina A, Dumonceau JM, Frossard JL. Proteomic Analysis of Human Bile from Malignant Biliary Stenosis Induced by Pancreatic Cancer Journal of Proteome Research.2009; 8(1):159-69
Guerrier L, Claverol S, Finzi L et al. Contribution of solid-phase hexapeptide ligand libraries to the repertoire of human bile proteins. Journal of Chromatography.2007;1176(1-2):192-205
Chen Bo, Zheng Jian-wei, Wang Jian-ming, et al. Establishment and preliminary analysis of a 2-D human biliary map Chinese Journal of Hepatobiliary Surgery.2007
Chen B, Dong JQ, Chen YJ et al Two-dimensional electrophoresis for comparative proteomic analysis of human bile. Hepatobiliary & pancreatic diseases international.2007 Aug;6(4):402-6
Guerrier L, Claverol S, Finzi L et al Contribution of solid-phase hexapeptide ligand libraries to the repertoire of human bile proteins.Journal of Chromatography A.2007;1176(1-2):192-205
Kristiansen TZ, Bunkenborg J, Gronborg M et al A Proteomic Analysis of Human Bile Molecular and Cellular Proteomics.2004;3:715-728
Organ Homogenates
Myerson, J., He, L., Lanza, G., Tollefsen, D. and Wickline, S. Thrombin-inhibiting perfluorocarbon nanoparticles provide a novel strategy for the treatment and magnetic resonance imaging of acute thrombosis. Journal of Thrombosis and Haemostasis.2011;9:1292-1300
Red Blood Cells
Antunes RF; Brandao C; Maia M; Arosa FA. Red blood cells release factors with growth and survival bioactivities for normal and leukemic T cells. Immunology and Cell Biology.2011;89(1):111-21
Tracheal Swab Samples
Li D, Wang J, Wang R, Li Y. A nanobeads amplified QCM immunosensor for the detection of avian influenza virus H5N1, Biosensors and Bioelectronics.2011;26(S10):4146-4154
Fu LM, Shinnick TM. Genome-wide exploration of the drug action of capreomycin on Mycobacterium tuberculosis using Affymetrix oligonucleotide GeneChips Journal of Infection.2007;54(S3):277-284
Fu LM, Shinnick TM. Genome-wide analysis of intergenic regions of mycobacterium tuberculosis H37Rv using affymetrix genechips. EURASIP journal on bioinformatics & systems biology.2007:23054
Tissue and Cell Culture
Alhamdani MS, Schroder C, Hoheisel JD. Analysis conditions for proteomic profiling of mammalian tissue and cell extracts with antibody microarrays. Proteomics.2010;10(17):3203-7
Czambel RK, Kharlamov A, Jones SC. Variations of brain endothelial nitric oxide synthase concentration in rat and mouse cortex.Nitric Oxide.2010;22(S1): 51-57
Plasma/Serum
Lijowski M, Caruthers S, Hu G. High-Resolution SPECT-CT/MR Molecular Imaging of Angiogenesis in the Vx2 Model Investigative Radiology.2009;44(1): 15–22
Turner JD, Langley RS, Johnston KL. Wolbachia Lipoprotein Stimulates Innate and Adaptive Immunity through Toll-like Receptors 2 and 6 to Induce Disease Manifestations of Filariasis The Journal of Biological Chemistry.2009;284:22364-22378
Torrelles JB, DesJardin LE, MacNeil J. et al Inactivation of Mycobacterium tuberculosis mannosyltransferase pimB reduces the cell wall lipoarabinomannan and lipomannan content and increases the rate of bacterial-induced human macrophage cell death Glycobiology.2009;19(7):743- 755
Cho N, Chueh PJ, Kim C et al Monoclonal antibody to a cancer-specific and drug-responsive hydroquinone (NADH) oxidase from the sera of cancer patients. Cancer Immunology, Immunotherapy. 2002;51(3):121-9
Shapiro S, Beenhouwer DO, Feldmesser M et al. Immunoglobulin G Monoclonal Antibodies to Cryptococcus neoformans Protect Mice Deficient in Complement Component C3 Infect. Infection and immunity.2002;70(5):2598-604
Castro AR, Morrill We, Pope V. Lipid Removal from Human Serum Samples Clinical and diagnostic laboratory immunology.2000;7(2):197-199
Nussbaum G, Cleare W, Casadevall A et al Epitope Location in the Cryptococcus neoformans Capsule Is a Determinant of Antibody Efficacy The Journal of experimental medicine.1997;185:685-694
Organ Homogenates
Thakuria D, Schmidt O, Liliensiek AK. Field preservation and DNA extraction methods for intestinal microbial diversity analysis in earthworms. Journal of Microbiological Methods.2009;76(3):226-33
Cheng AM, Moore EE, Masuno T et al Normal Mesenteric Lymph Blunts the Pulmonary Inflammatory Response to Endotoxin. Journal of Surgical Research.2006;136(S2):166-171
McNally T, Mackie IJ, Machin SJ et al. Increased levels of beta 2 glycoprotein I antigen and beta 2 glycoprotein I binding antibodies are associated with a history of thromboembolic complications in patients with SLE and primary antiphospholipid syndrome British journal of rheumatology.1995 Nov;34(11):1031-6
Saliva
Garton NJ, Waddell SJ, Sherratt Al et al. Cytological and Transcript Analyses Reveal Fat and Lazy Persister-Like Bacilli in Tuberculous Sputum.PLoS medicine.5(4): e75
Lucy E. DesJardin Isolation of M. tuberculosis RNA from Sputum Methods in Molecular Medicine.2001;48:133-139
Beenhouwer DO, Shapiro S, Feldmesser M et al. Both Th1 and Th2 Cytokines Affect the Ability of Monoclonal Antibodies To Protect Mice against Cryptococcus neoformansInfection and immunity.2001;69: 6445-6455
Desjardin LE, Perkins MD, Wolski K et al. Measurement of Sputum Mycobacterium tuberculosis Messenger RNA as a Surrogate for Response to Chemotherapy American journal of respiratory and critical care medicine.1999;160(1):203-10
Template Preparation Production Sequencing Protocols. Stanford Genome Technology Center Patents
J Krupey - United States Patent: 5885921 Hydrophobic silica adsorbents for lipids
David C. Jones. United States Patent: 7999084. Devices and methods for reducing matrix effects DJ Morre, NM McCarty, D Morre et al United States Patent: 7053188. Monoclonal antibodies specific for neoplasia-specific NADH: disulfide reductase
Morre, James D et al. United States Patent: 20030170757. Monoclonal antibodies specific for neoplasia-specific NADH: disulfide reductase
Mcintyre, John A. United States Patent: 20120107841. Serum Diagnostic Method, Biomarker and Kit for Early Detection and Staging of Alzheimer's Disease
CONTACT US
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