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 Price USD$285.00 SKU 17400 Format Micro Midi Maxi Size 25 Preps Quantity 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 PI17400-13 ProteoSpin Urine Protein Concentration Micro Kit Insert.pdf PI52300-4 Proteospin Urine Protein Concentration Kit Insert.pdf PI21600-12 ProteoSpin™ Urine Protein Concentration Maxi Kit Insert .pdf SAFETY DATA SHEETS 17400_21600_52300_SDS.pdf POSTERS Poster 28 - Proteomic Profiling of Prostate Cancer and Benign Prostatic Hyperplasia from Urine.pdf Poster 11 - Urine Proteomic Pattern Analysis For Early Hepatitis and Hepatitis related Liver Cancer Detection-min.pdf RELATED BLOGS 3 Reasons why Norgen's Industry-Leading Patented RNA Purification Technology is Best-in-Class Citations Powered by Bioz See more details on Bioz 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 Price USD$285.00 SKU 17400 Format Micro Midi Maxi Size 25 Preps Quantity