Stain proteins in 1D and 2D gels with this rapid, ultra-sensitive and versatile silver stain system that yields consistent and reliable results. Wash gel with water. The Pierce Silver Stain Kit is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background while delivering consistent, high-sensitivity staining results at each and every use. LC3B puncta and autophagic flux assay. Destain Reagent B, 14mL . In standard mini gels, proteins are detectable at greater than 0.25ng per band or spot. Reversibly stain proteins in polyacrylamide gels to identify bands to excise; then destain the gel pieces for analysis by mass spectrometry. 4-20% Precise Gel containing two-fold serial dilutions of purified proteins. For analysis on one of our lower sensitivity instruments, the cost is $82 per band for Coomassie stain and $92 for Silver Stain. GE Healthcare plusone dna silver staining kit Plusone Dna Silver Staining Kit, supplied by GE Healthcare, used in various techniques. However, the molecular mechanism of action of IVM on A. simplex s.s. remains unknown. Bands should consist of no more than 10mm length of a single lane from a gel. Shop Products Antibodies & Protein Biology. Pierce Silver Stain Kit for Mass Spectrometry (24600)) or reversible zinc staining (Pierce Zinc Reversible Stain Kit (24582)), alternative destaining procedures will be required. 4.1.1 Save a 1 g aliquot of the lysate at this step and freeze it at -80C for use as the "input" fraction for quality control by SDS-PAGE and silver staining. Conveniently packaged, extremely stable, and highly sensitive, ProteoSilver is an ideal product for any protein scientist. English Deutsch; Change Country. Many of the fluorescent stains are also compatible with mass spectrometry. The kit contains prepared solutions of silver staining reagents along with detailed instructions to achieve optimal results. A Pierce BCA protein assay kit (Thermo Fisher Scientific) was used to detect the total protein concentration. Pierce Silver Staining for Mass Spectrometry Kit, Thermo #24600) 3. Commercial silver staining kits compatible with mass. Silver staining and destain of separated proteins was performed per manufactures instructions using the Pierce silver stain kit for mass spectrometry (Thermo Fisher Scientific, Waltham MA). 2 Thermo Scientific Pierce Color Silver Stain Kit Detect proteins with this metallic silver staining technique that produces color variations to differentiate protein bands and spots in 1D and 2D gels. For silver staining, blots were developed with the Pierce Silver Stain for Mass Spectrometry Kit (Pierce Cat# 24600). Mouse brain tissue (0.25 g) was homogenized with a tissue tearer and the proteins were extracted using the Thermo Scientific Pierce Mass Spec Sample Prep Kit for Cultured Cells. The Penn State College of Medicine Mass Spectrometry and Proteomics Core provides mass spectrometry analyses including identification and quantitation of proteins, peptides, oligonucleotides, carbohydrates and small molecules like drugs, metabolites, and lipids. Rating: 1 (959 Rating) Highest rating: 4. Silver staining was performed using Pierce Silver Stain for Mass Spectrometry kit (Catalog#:24600) according to the manufacturer's protocol. Lowest rating: 1. Silver staining is not recommended for agarose gels. Specifications Detection Location In-Gel Detection "Many silver staining protocols and commercial staining kits are not compatible with mass spectrometry due to the use of crosslinking reagents. 24612). Proteins identified by mass spectrometry. To identify proteins isolated from . For western blotting, proteins were transferred to a PVDF membrane (Millipore), blocked in 4% milk in TBST (0.05 M Tris pH7.2, 0.15 M NaCl, 0.1% Tween20) for 1 h at room temperature and incubated with primary antibodies . No offer available . pierce silver stain kit for mass spectrometry. The improved SilverSNAP Stain Kit II is an easy and fast way to detect proteins of low concentration in polyacrylamide gels. 2.10 Immunoblot and coimmunoprecipitation assay. Catalog Number: (PI24600) Supplier: . . Pierce Silver Stain for Mass Spectrometry, Thermo Scientific. I followed the Pierce silver stain kit (Thermo fisher #24612) protocol. described in this Tech Tip has been validated for use with the Pierce Silver Stain Kit. It is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background. Descriptions: The Pierce Silver Stain Kit is a rapid, ultra-sensitive, and versatile silver stain system for protein detection in polyacrylamide gels, yielding consistent . For best results, use the Pierce Silver Stain Kit for Mass Spectrometry (Product No. Please contact Laura Bond directly at lbond@boisestate.edu or 208-426-1438. Antibodies & Protein Biology H 2 O SS4. For Research Use Only. The excised pieces were destained using the reagents and procedure provided in the Pierce Silver Stain for Mass Spectrometry kit. Silver staining was performed using Pierce Silver Stain Kit (Thermo Fisher,24600) according to the manufacturer's protocol. Figure 2. . The Pierce Silver Stain Kit is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background while delivering consistent, high-sensitivity staining results at each and every use. Pierce Silver Stain Kit For Mass Spectrometry 30-31 Pierce Silver Stain II 32 Pierce Color Silver Stain 33 Pierce Silver Stain Rescue Reagent 34 Pierce Zinc Reversible Stain 35 Pierce Glycoprotein Stain 36 Krypton Glycoprotein Staining Kit 37 GelCode 6xHis Protein Tag Staining Kit 38 GelCode Phosphoprotein Staining Kit 39 . ALP activity in each sample was measured as units per gram of protein per 15 minutes. Thermo Scientific Pierce Silver Stain for Mass Spectrometry Reversibly stain proteins in polyacrylamide gels to identify bands to excise; then destain the gel pieces for analysis by mass spectrometry. Acrylamide modification of cystine results in a peptide mass increase of 71.04. PI24600 $349.00 / Each of 1 Qty Check Availability Add to cart Description Specifications Silver dye stains Silver staining is the most sensitive colorimetric method for detecting total protein. Retrieving Each-+ Pig Trypsin. Life scientists around the world look to PierceStains for speed, ease of use, sensitivity, superior results and economy. Ivermectin (IVM), an antiparasitic drug, has a positive effect against Anisakis simplex s.s. infection and has been used for the treatment and prevention of anisakiasis in humans. 1.Pierce Silver Stain Kit - Thermo Fisher Scientific. The gels were then either stained with Colloidal Coomassie Blue (GelCode Blue Safe Protein StainThermo Scientific, Pierce, Rockford, IL, USA) or with Pierce Silver Stain Kit for Mass Spectrometry (Thermo Scientific). Fix gel in Fixing Solution3. Life Technologies SilverQuest Pierce Silver Stain Kit Sigma ProteoSilver. Bioz Stars score: 95/100, based on 19 PubMed citations. Pierce Silver Stain for Mass Spectrometry 1 L 24600 Pierce Zinc Reversible Stain Kit 1.5 L 24582 PageRuler Unstained Protein Ladder 2 x 250 L 26614 For protein bands stained with mass spectrometry-compatible silver stains (e.g. Silver. It is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background. Thermo Scientific Pierce Silver Stain for Mass Spectrometry Reversibly stain proteins in polyacrylamide gels to identify bands to excise; then destain the gel pieces for analysis by mass spectrometry. High-purity - mass spectrometry-grade reagents in a non-leachable . However, this and other standard silver staining methods are not compatible with mass spectrometry (MS), which is fast . Cost of services will be made available by quote after consultation. Workflow-compatible - mild chemical formulation ensures compatibility with mass spectrometry, sequencing and other downstream . Thermo Scientific Pierce Silver Stain Kit is a rapid, ultra-sensitive and versatile silver stain system for protein detection in polyacrylamide gels, yielding consistent and reliable results with many 1D and 2D gel types for both first-time and experienced users. Extract the gel by adding 50 l 0.1% TFA, 60% CH3CN and shaking at room temperature for 60 minutes (use more volume if necessary) Repeat steps 8 and 9, twice and Combine extracts with the supernatant from step 8. The whole page was stained by Pierce Silver Stain for Mass Spectrometry Kit (Thermo Scientific) and evaluated with mass spectrometry assay. We prefer Coomassie blue-visible protein bands, in case of silver staining use an MS compatible silver staining method (e.g. . Thermo Scientic Pierce Gel Stains provide speed, simplicity, convenience and economy over both homemade and other commercially available staining products. Gently agitate the gel in the staining solution for an additional 20 minutes. PI24600 $448.83 / Each Quantity Add to cart Description Specifications Documents Description It is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background. Our mass spectrometry analysis identified a list of presumptive endothelial PKA substrates, which will potentially also be involved in angiogenesis. Stain, 500mL . Negative staining techniques such as zinc stain are compatible with mass spectrometry. 4 Thermo Scientific Pierce Silver Stain Rescue Reagent After silver staining, bands were excised and analyzed by mass spectrometry, which showed that hnRNP K interacts with . This strip will function as the "reference" gel strip. Briefly, 10 mM DTT was applied at 56 C for reduction of disulfide bonds and 55 mM IAM at room temperature in the dark . The major P. sativum . Storage: Upon receipt store at ambient temperature. 24600), which includes an optimized procedure and all the necessary reagents for staining gels and destaining gel pieces before in-gel trypsin digestion and MS. 2.12 RNA pull-down assay and mass spectrometry. In standard mini gels, proteins are detectable at greater than 0.25ng per band or spot. Promotions are available 18 Hoefer TE22 Mini Tank Blotting Unit Safe, economical unit requires minimal current and only about 1L of buffer 19 3 Thermo Scientific Pierce Color Silver Stain Kit Detect proteins with this metallic silver staining technique that produces color variations to differentiate protein bands and spots in 1D and 2D gels. 6.2 Silver staining after Coomassie Brilliant Blue R-250 staining 6.1 Nucleic acid staining Follow the silver staining protocol, deleting the acetic acid from the fixatives (steps 1, 2, and 3, Table 1). Supplier: Thermo Scientific The Pierce Silver Stain Kit is a rapid, ultra-sensitive, and versatile silver stain system for protein detection in polyacrylamide gels, yielding consistent and reliable results. Silver. Discard staining solution and rinse the gel two times with . Pierce Silver Stain uses formaldehyde in the stain and developer working solutions. The Thermo Scientific Pierce Silver Stain Kit is a rapid, ultra-sensitive and versatile silver stain system for protein detection in polyacrylamide gels, yielding. Silver. Gel cuts are submitted in a tube and covered by 3% acetic acid in ultra-pure water. With a detection limit of 0.1 ng/mm2 of protein (BSA) and an extremely low background . The Pierce Silver Stain Kit is a rapid, ultra-sensitive and versatile silver stain system for protein detection in polyacrylamide gels, yielding consistent and reliable results with many 1D and 2D gel types Thermo Scientific Pierce Silver Stain Kit The supplier does not provide quotations for this particular product through SelectScience. We offer several optimized silver staining kits depending on your research needs. Silver ions (from silver nitrate in the staining reagent) interact and bind with certain protein functional groups. Protein Quantitation and Staining 36-42 Introduction 36 BCA Protein Assay 37 Micro BCA Assay 38 Silver Stain for Mass Spectrometry 39 Protein Stains 41 Protein Digestion 43-53 Introduction 43 Trypsin Protease, MS Grade 44 LysN Protease, MS Grade 46 Lys-C Protease, MS Grade 47 Asp-N Protease, MS Grade 47 Glu-C Protease, MS Grade 48 SPECIAL INTERESTS 1 - 30 of 70 Results 1 Thermo Scientific Pierce Silver Stain Kit Stain proteins in 1D and 2D gels with this rapid, ultra-sensitive and versatile silver stain system that yields consistent and reliable results. The specific strip was cut for mass spectrometry. However, the procedure accompanying the kit for mass spectrometry is optimized to maximize polypeptide recovery without greatly sacrificing sensitivity. 3 Introduction The Thermo Scientific Pierce Silver Stain for Mass Spectrometry is a complete kit for rapid and ultra-sensitive silver staining . In addition, BioS1 in-solution tryptic digests were separated using Tricine gels (Invitrogen). (SDS-PAGE) and stained with Pierce Silver Stain Kit (24612, Thermo Fisher Scientific) according to the instruction manual. The Pierce Silver Stain Kit is a rapid, ultra-sensitive, and versatile silver stain system for protein detection in polyacrylamide gels, yielding consistent and reliable results. experiment was performed using a Pierce Co-IP Kit (Thermo Fisher . In comparison to the former Silver SNAP Kit, the new kit was improved to reduce time and enhance flexibility: the washing times are reduced and the fixing and staining steps can be extended overnight. Manufacturer: Thermo Scientific 0024600 Catalog No. Related Questions 2 Invitrogen SimplyBlue SafeStain Less hazardous and does not require methanol or acetic acid fixatives or destains 3 Not for use in diagnostic procedures. Publish: 13 days ago. Protein concentrations in the . Learn more about the different protein gel and membrane staining methods from Coomassie, silver, fluorescent and functional group stains. Antibodies & Protein Biology Antibody Production & Purification; Electrophoresis, Western Blotting and ELISA Destain Reagent A, 4mL . Tech Tip #50: Process stained polyacrylamide gels for mass spectrometry B. Silver staining with sodium dodecyl sulfate polyacrylamide gel electrophoresis gels was performed by using Pierce Silver Stain for Mass Spectrometry Kits. Mass Spectrometry. 2. Best practice is freeze at -80 > 3hr and then lyophilize. For Research Use Only. Available support is determined case-by-case. The technique involves the deposition of metallic silver onto the surface of a gel at the locations of protein bands. Stain Enhancer, 25mL . The Pierce Silver Stain Kit is a rapid, ultra-sensitive, and versatile silver stain system for protein detection in polyacrylamide gels, yielding cons . Manufacturer: Thermo Scientific 24600 Catalog No. Certificates. For analysis on our highest sensitivity instrument, the cost is $204 per band for Coomassie stain and $224 for Silver Stain. Decant Sensitizing Solution and rinse5. Antisera . September 9, 2022 . Description: Pierce Silver Stain for Mass Spectrometry is a complete kit for rapid and ultra-sensitive silver staining of proteins in polyacrylamide gels and effic. Herein, tandem mass tag (TMT) labeling and extensive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS . SDS-PAGE was used to separate the pull-down proteins, and silver staining was subsequently performed using Pierce Silver Stain for Mass . Do not allow the reagent to boil. Align the stained strip with the unstained gel portion, . . Shop. Thermo Scientific Pierce Silver Stain for Mass Spectrometry. What Does it Cost? No. . The Pierce Silver Stain Kit is a rapid, ultra-sensitive, and versatile silver stain system for protein detection in polyacrylamide gels, yielding consistent and reliable results. Catalog Number: (PI24600) Supplier: Thermo Scientific. . For immunoblot analysis, cells or tissues were lysed with NP-40 lysis buffer supplemented with a protease inhibitor cocktail. The Pierce Silver Stain Kit is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background while delivering consistent, high-sensitivity staining results at each and every use. Gel bands from which proteins were identified are indicated at the left. Tissue lysate (100 g) was subjected to sample preparation workflow and sample (500 ng) was analyzed by LC-MS/MS on a Thermo Scientific Velos Pro ion trap mass . The following in-gel digestion was conducted according to the protocol of Shevchenko et al. Creation of TGD2 antiserum was described previously . Supplier: G . Double and single stranded DNA and RNA in poly-acrylamide gels can be stained. Figure 39. . Compare different protein staining methods Highlights of silver stains: Not for use in diagnostic procedures. Gels were immunoblotted or stained with the Pierce Silver Stain kit for mass spectrometry (Thermo Fisher Scientific) according to instructions. If we are unable to provide services, we may be able to help find you the appropriate support through our network. Silver Stain Silver staining is a more sensitive, but more difficult, method for visualizing protein bands in . Which stains (other than Coomassie blue) are compatible with mass spectrometry? Example gels stained with the Pierce Silver Stain Kit Figure 1. Stain Developer, 500mL . However, the procedure accompanying the kit for mass spectrometry is optimized to maximize polypeptide recovery without greatly sacrificing sensitivity. . Get All The Latest News Help & Support Center English Deutsch; Change Country; 0. Specifications Detection Location In-Gel Detection In addition, 20 L of these elution samples (high or low zonulin) were separated by SDS-PAGE using precast Bolt 4-12% Bis-Tris Plus gels (ThermoFisher, Waltham, MA, USA). gel 2 times with ultrapure water. Trypsin-digested protein samples were analyzed with LTQ-Orbitrap XL (Thermo Fisher Scientific) mass spectrometry. Author: www.thermofisher.com. Pierce Silver Stain uses formaldehyde in the stain and developer working solutions. Pierce Silver Stain Kit for Mass Spectrometry 30-31 Pierce Silver Stain II 32 Pierce Color Silver Stain 33 Pierce Silver Stain Rescue Reagent 34 Pierce Zinc Reversible Stain 35 Pierce Glycoprotein Stain 36 Krypton Glycoprotein Staining Kit 37 GelCode 6xHis Protein Tag Staining Kit 38 GelCode Phosphoprotein Staining Kit 39 . Thermo Scientific Pierce Silver Stain for Mass Spectrometry Reversibly stain proteins in polyacrylamide gels to identify bands to excise; then destain the gel pieces for analysis by mass spectrometry. 3. Silver. 4.2 Add washed beads (prepared as above) to the samples in a ratio of 5-10 g of beads to 1 g of protein and briefly vortex. 4. incubate gel in 2 changes of Sensitizing Solution. Decant Fixing Solution and for 10 minutes. Silver. ZERO BIAS - scores, article reviews, protocol conditions and more If mass spectrometry is the intended downstream application, choose one of our two silver stain kits optimized for peptide recovery after in-gel trypsin digestion for mass spectrometry. Pierce Silver Stain for Mass Spectrometry, Thermo Scientific. Recommended SDS PAGE Stain Protocols Kits like GelCode Blue from Pierce and Biosafe Coomassie from Biorad are NOT compatible for in-gel digestion and mass spectrometry analysis unless you do a fixing step first. Proteins were stained using the Pierce silver stain for mass spectrometry (ThermoFisher) or detected by Western Blot. Additional Recharge Centers Recharge Centers The third band from the top is BSA, with the rightmost lane containing 14ng. Related Pierce Products 24612 SilverSNAP Stain Kit II 24600 SilverSNAP Stain for Mass Spectrometry 24597 Color Silver Stain Kit 24590 GelCode Blue Stain Reagent, 500 ml, coomassie G-250 stain 24615 Imperial Protein Stain, 1 L, coomassie R-250 stain 24582 E-Zinc Reversible Stain Kit the Thermo Scientific Pierce Silver Stain Kit (Cat. . Discard the last wash and add 20mL of PageBlue Protein Staining Solution, or sufficient volume to cover the gel, and microwave on high power for 20-30 seconds. In standard mini gels, proteins are detectable at greater than 0.25ng per band or spot. 2 -D gel of 15g of TNF Incubate gel in Staining Solution. . Briefly, the SDS-page gel was washed in ultrapure water and fixed . The Pierce Silver Stain Kit is a metallic silver (Ag) protein stain that yields a remarkably clear and uniform gel background while delivering consistent, high-sensitivity staining results at each and every use. (You should see no powder [detergent] or salt). 2.