ProteoSpin™ Urine Protein Concentration Kits For the rapid and efficient concentration of total proteins from urine samples For research use only and NOT intended for in vitro diagnostics. Powered by Bioz ProteoSpin™ Urine Protein Concentration Kits For the rapid and efficient concentration of total proteins from urine samples SKU 17400 Format Micro Midi Maxi Size 25 Preps Contact Distributor Overview Details Documentation Citations Overview Simultaneous concentration, desalting and buffer exchange of total urinary proteins No molecular weight cutoff allows for isolation of all sizes of proteins and peptides Purification is based on spin column chromatography that uses Norgen’s resin separation matrix These kits concentrate urine proteins while simultaneously removing salts, urea, and other urine contaminants. There is no molecular weight cut-off and therefore the columns capture total urinary proteins and peptides of all sizes making them ideal for biomarker discovery work, differential expression of proteins in various diseases, or other diagnostic research. The columns are convenient, rapid and easy to use and thus offer significant time savings over classic dialysis protocols. The resulting high-quality protein sample is concentrated and free from the original sample salts, thus preparing the sample conveniently for downstream proteomic applications including SDS-PAGE, 2D Gels, MALDI-TOF, LC/MS, LC/MS/MS, whole protein mass spectrometry, western blotting, protein microarrays, and more. ProteoSpin™ Urine Protein Concentration Micro Kit Each spin column is able to concentrate and desalt up to 200 μg of urine proteins. Twelve samples can be processed in 20 minutes. ProteoSpin™ Urine Protein Concentration Midi Kit The ProteoSpin™ Urine Protein Concentration Midi Kit provides a fast and simple procedure for concentrating dilute solutions of urine proteins from 1 to 5 mL inputs of urine. Each mini spin column is able to concentrate and desalt up to 3 mg of urine proteins in 30 minutes. ProteoSpin™ Urine Protein Concentration Maxi Kit The ProteoSpin™ Urine Protein Concentration Maxi Kit provides a fast and simple procedure for concentrating dilute solutions of urine proteins from 2 to 20 mL inputs of urine. Each maxi spin column is able to concentrate and desalt up to 4 mg of urine proteins in 45 minutes. Details Supporting Data Previous Figure 1. Two-Dimensional Gel Analysis. Total urinary proteins were purified from 1 mL urine samples collected from a Hepatitis B virus infected patient as well as a normal healthy individual using this kit. The protein samples were then separated on IPG strips (4-7) for 2D SDS-PAGE analysis. A total of 30 μg protein was loaded to the ReadyStrip™ IPG strip pH 4-7 and focused on a PROTEAN IEF Cell at 250 V for 20 minutes, followed by 4000 V for 1 hour and then to 20,000 V. The strips were then transferred to the second dimension where they were loaded onto a 12% SDS-PAGE and run at 200 V for 75 minutes. The gels were stained using the Silver Stain Plus Kit from Bio-Rad and pictures were taken using an AlphaImager™ 2200 from AlphaInnotech. The protein profiles of the Hepatitis B Virus infected patient and the normal individual (Control) are displayed above. The control 2D gel and the HBV 2D gel were analyzed and compared using Progenesis SameSpots software from Nonlinear Dynamics. The two gels were wrapped and overlaid to show the common spots as well as the difference between the two gels. The results suggested that this purification method is completely applicable for 2D protein analysis. Figure 2. Consistent Protein Purification from 1 mL Urine Samples. Seven different urine samples were collected, and 1 mL from each sample was processed using the Urine Protein Concentration Micro Kit according to the bind, wash and elute procedure. The concentrated urinary proteins were eluted in 100 µL of Elution Buffer, and 10 µL of each elution was resolved on a 12% SDS-PAGE gel and run at 200 V for 75 minutes. The gels were stained using Commassie Blue and pictures were taken using an AlphaImagerTM 2200 from AlphaInnotech. As it can be seen, consistent protein recovery was observed in each case. Figure 3. Western Blot Showing the Recovery of Free PSA, ACT-bound PSA and a mixture of both from Spiked Urine Samples Detected using a Combination of Anti-free PSA Antibodies and Anti-ACT-bound PSA Antibodies. Five milliliter urine samples were spiked with free-PSA, ACT-bound PSA and a combination of free-PSA and ACT-bound PSA. Total urinary proteins, including the spiked-PSA, were then purified using Norgen’s Urine Protein Concentration Kit. The urinary proteins were concentrated in 120 µL of Elution Buffer, and 12 µL of each elution was loaded onto a native 12% PAGE gel and run at 200 V for 75 minutes. Next, the proteins were transferred to a PVDF membrane using standard laboratory procedures. Once transferred, the membrane was probed using the Santa Cruz Anti-PSA antibody against free PSA and the Santa Cruz Anti-PSA/ACT antibody which is highly specific against ACT-bound PSA. The Sigma Anti-Mouse IgG-HRP antibody was used as the secondary antibody. The membrane was then developed using a Pierce CN/DAB Kit. In the gel above, Lane 1 corresponds to free PSA, Lane 2 corresponds to ACT-bound PSA, Lane 3 corresponds to the free PSA and ACT-bound PSA mixture, Lane C corresponds to the input of both the free PSA and the ACT-bound PSA, and Lane M is the marker lane. Figure 4. High Yield of Urine Proteins. Five different urine sample volumes were concentrated using Norgen’s Urine Protein Concentration Midi Kit. The total urine protein yield from the different urine sample volumes processed using Norgen’s kit appears to be increasing with the increase of the urine sample volume. Figure 5. SDS-PAGE Gel Showing Total Proteins Purified from Different Urine Sample volumes using Norgen’s Urine Protein Concentration Midi Kit. Five different urine sample volumes were processed using the Urine Protein Concentration Midi Kit according to the bind, wash and elute procedure. The concentrated urinary proteins were eluted in the volumes recommended in the protocol based on the input amount, and 10 µL of each elution was resolved on a 12% SDS-PAGE gel and run at 200 V for 75 minutes. The gels were stained using Commassie Blue and pictures were taken using an AlphaImager™ 2200 from AlphaInnotech. As it can be seen, total protein yield increases with the increase of the urine sample volume. Figure 6. No Loss of Urine Proteins. The urinary proteins present in the 20 mL urine sample were concentrated into 2 mL of the provided Elution Buffer. The urinary proteins were resolved on a 12% SDS-PAGE gel run at 200 V for 75 minutes, and were stained used Commassie Blue staining. In the gel, Lane 1 contains 25 µL of input urine before concentration, Lane 2 contains 15 µL of the binding flowthrough, and Lane 3 contains 15 µL of the concentrated eluted protein. No urine proteins are lost during the binding step, as can be seen by examining the binding flowthrough. Figure 7. Concentration of Urine Proteins. The urinary proteins present in 20 mL of urine were concentrated into 2 mL of Elution Buffer using Norgen’s Urine Protein Concentration Maxi Kit according to the provided bind, wash and elute procedure. The urinary proteins were resolved on a 12% SDS-PAGE gel run at 200 V for 75 minutes, and the urinary proteins were stained used Commassie Blue staining. Lane 1 contains 20 µL of the urine sample that was used as the input, Lane 2 contains 5 µL of the elution, Lane 3 contains 10 µL of the elution, and Lane 4 contains 20 µL of the elution. By comparing Lanes 1 and 4, which contain equal volume amounts of input and output, it can clearly be seen that the urine proteins have been highly concentrated though the use of the kit. Next Click for expanded view Kit Specifications Maximum Urine Input Volume 1 mL Maximum Recovered Protein 200 μg Elution Volume 100 μL Time to Process 12 Samples 20 minutes Storage Conditions All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 2 years after the date of shipment. Component Cat. 17400 (25 preps) Cat. 52300 (10 preps) Cat. 21600 (4 preps) Wash Solution C 60 mL 60 mL 130 mL Binding Buffer A 4 mL 4 mL 8 mL Elution Buffer C 8 mL 30 mL 30 mL Protein Neutralizer 4 mL 4 mL 4 mL Micro Spin Columns 25 - - Midi Spin Columns (assembled with collection tubes) - 10 - Maxi Spin Columns (assembled with collection tubes) - - 4 Collection Tubes 25 - - Elution Tubes (1.7 mL) 25 - - Midi Elution Tubes - 10 - Elution Tubes (50 mL) - - 4 Product Insert 1 1 1 Documentation PROTOCOLS (17400) ProteoSpin™ Urine Protein Concentration Micro Kit - Protocol (25 preps) (52300) ProteoSpin™ Urine Protein Concentration Midi Kit - Protocol (10 preps) (21600) ProteoSpin™ Urine Protein Concentration Maxi Kit - Protocol (4 preps) SAFETY DATA SHEETS (17400) ProteoSpin™ Urine Protein Concentration Micro Kit - Safety Data Sheet POSTERS Urine Protein Concentration Micro Kit Proteomic Profiling of Prostate Cancer and Benign Prostatic Hyperplasia From Urine Present Urine Protein Concentration Micro Kit The Urinary Genomic and Proteomic Profiling of Hepatocellular Carcinoma Patients Infected RELATED BLOGS 3 Reasons why Norgen's Industry-Leading Patented RNA Purification Technology is Best-in-Class Citations Micro (17400) Title Peptidomics analysis reveals changes in small urinary peptides in patients with interstitial cystitis/bladder pain syndrome Journal Scientific Reports. 2022. Authors Md Shadman Ridwan Abid, Haowen Qiu, Bridget A. Tripp, Aline de Lima Leite, Heidi E. Roth, Jiri Adamec, Robert Powers & James W. Checco Title KeepEX, a simple dilution protocol for improving extracellular vesicle yields from urine Journal European Journal of Pharmaceutical Sciences. 2016. Authors Puhka M, Nordberg ME, Valkonen S, Rannikko A, Kallioniemi O, Siljander P, Hällström TA Title Mass Spectrometric Quantification of Urinary Netrin-1 Protein in Renal Transplant Recipients Journal Current Proteomics. 2016. Authors Ozcan, Filiz; Akbas, Halide; Suleymanlar, Gultekin; Aslan, Mutay; Yucel, Gultekin Title Cisplatin nephrotoxicity in male beagle dogs: next-generation protein kidney safety biomarker tissue expression and related changes in urine Journal Toxicology Research. 2016. Authors J. E. McDuffie, Y. Chen, J. Y. Ma, S. Lee, K. M. Lynch, D. M. Hamlin, L. Nguyen, M. Rizzolio, M. Sonee and S. Snook Title Mass spectrometric quantification of urinary human liver fatty acid binding protein in renal transplant recipients Journal Rapid Communications in Mass Spectrometry. 2016. Authors Filiz Ozcan, Halide Akbas, Ebru Kırac, Gultekin Suleymanlar, Mutay Aslan, andGultekin Yucel Title Comparison of Depletion Strategies for the Enrichment of Low-Abundance Proteins in Urine Journal PLOS ONE. 2015. Authors Szymon Filip, Konstantinos Vougas, Jerome Zoidakis, Agnieszka Latosinska, William Mullen, Goce Spasovski, Harald Mischak, Antonia Vlahou, Joachim Jankowski Title Potential Urinary Protein Biomarker Candidates for the Accurate Detection of Prostate Cancer among Benign Prostatic Hyperplasia Patients Journal Journal of Cancer. 2014. Authors Taha A Haj-Ahmad, Moemen AK Abdalla, Yousef Haj-Ahmad Title Promising Candidate Urinary MicroRNA Biomarkers for the Early Detection of Hepatocellular Carcinoma among High-Risk Hepatitis C Virus Egyptian Patients. Journal Journal of Cancer. 2012. Authors Abdalla M, Haj-Ahmad Y. Title Human Alpha Defensin 5 Expression in the Human Kidney and Urinary Tract. Journal PLOS One. 2012. Authors Spencer J, Hains D, Porter E, Bevins C, DiRosario J, Becknell B, Wang H, Schwaderer A. Title Ribonuclease 7 is a potent antimicrobial peptide within the human urinary tract. Journal Kidney International. 2011. Authors Spencer JD, Schwaderer AL, Dirosario JD, McHugh KM, McGillivary G, Justice SS, Carpenter AR, Baker PB, Harder J, Hains DS. Title Plasmin activates epithelial Na+ channels by cleaving the gamma subunit. Journal Journal of Biological Chemistry. 2008. Authors Lu CY, Chang YM, Tseng WL, Feng CH, Lu CY. Title Large-scale protein identification of human urine proteome by multi-dimensional LC and MS/MS. Journal Proteomics Clinical Applications. 2007. Authors Chen YT, Tsao CY, Li JM, Tsai CY, Chiu SF, Tseng TL. Maxi (21600) Title Treatment of Collagen Defects Using Protein Solutions Journal United States Patent Application 20160166645. 2016. Authors Matuska A, O'shaughnessey K, Woodell-may JE Title Methods of Treating Pain Using Protein Solutions Journal United States Patent Application 20160136245. 2016. Authors Toler K, King W, Woodell-may JE Title Profiling of urinary proteins in Karan Fries cows reveals more than 1550 proteins. Journal Journal of Proteomics. 2015. Authors Bathla S, Rawat P, Baithalu R, Yadav ML, Naru J, Tiwari A, Kumar S, Balhara AK, Singh S, Chaudhary S, Kumar R, Lotfan M, Behare P, Phulia SK, Mohanty TK, Kaushik JK, Nallapeta S, Singh I, Ambatipudi SK, Mohanty AK Title Plasmin activates epithelial Na+ channels by cleaving the gamma subunit. Journal Journal of Biological Chemistry. 2008. Authors Lu CY, Chang YM, Tseng WL, Feng CH, Lu CY. Title Large-scale protein identification of human urine proteome by multi-dimensional LC and MS/MS. Journal Proteomics Clinical Applications. 2007. Authors Chen YT, Tsao CY, Li JM, Tsai CY, Chiu SF, Tseng TL. Related Products Urine DNA Isolation Kit (Slurry Format)
ProteoSpin™ Urine Protein Concentration Kits For the rapid and efficient concentration of total proteins from urine samples For research use only and NOT intended for in vitro diagnostics. Powered by Bioz ProteoSpin™ Urine Protein Concentration Kits For the rapid and efficient concentration of total proteins from urine samples SKU 17400 Format Micro Midi Maxi Size 25 Preps Contact Distributor