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Ready-To-Run Buffers and Solutions
Electrophoresis Ready-to-Run Buffers and Solutions Bio-Rad is a premier provider of buffers and premixed reagents for life science research. We offer a variety of different products for all your protein and nucleic acid experiments. Whether you need powdered reagents or premixed solutions, Bio-Rad reagents meet the highest quality standards to ensure consistency and reliability in your experiments. FastCast™ Solutions FastCast Solutions Access all the benefits of TGX™ and Solution Applications TGX Stain-Free™ gel chemistries with TGX™ FastCast™ acrylamide starter kit, 7.5% Protein Separation the fastest gel run times, the most TGX FastCast acrylamide kit, 7.5% Protein Separation efficient transfers, stain-free visualization TGX FastCast acrylamide starter kit, 10% Protein Separation in 5 minutes, as well as a long shelf life TGX FastCast acrylamide kit, 10% Protein Separation (1 month after casting) using convenient TGX FastCast acrylamide starter kit, 12% Protein Separation premixed handcasting solutions. TGX FastCast acrylamide kit, 12% Protein Separation Long shelf life TGX chemistry retains TGX Stain-Free™ FastCast™ acrylamide starter kit, 7.5% Protein Separation Laemmli-like separation characteristics TGX Stain-Free FastCast acrylamide kit, 7.5% Protein Separation while using a standard Tris/glycine TGX Stain-Free FastCast acrylamide starter kit, 10% Protein Separation running buffer system. TGX Stain-Free FastCast acrylamide kit, 10% Protein Separation TGX Stain-Free FastCast acrylamide starter kit, 12% Protein Separation TGX Stain-Free FastCast acrylamide kit, 12% Protein Separation Gel Casting Solutions Gel Casting Buffers Tris Buffers and Acrylamide Solutions Solution Applications for Gel Casting 1.5 M Tris-HCI, pH 8.8 Resolving gel preparation Bio-Rad offers a variety of prepared 0.5 M Tris-HCI, pH 6.8 Stacking gel preparation solutions for casting polyacrylamide Acrylamide Solutions gels. -
Thermodynamic Quantities for the Ionization Reactions of Buffers
Thermodynamic Quantities for the Ionization Reactions of Buffers Robert N. Goldberg,a… Nand Kishore,b… and Rebecca M. Lennen Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899 ͑Received 21 June 2001; accepted 16 September 2001; published 24 April 2002͒ This review contains selected values of thermodynamic quantities for the aqueous ionization reactions of 64 buffers, many of which are used in biological research. Since the aim is to be able to predict values of the ionization constant at temperatures not too far from ambient, the thermodynamic quantities which are tabulated are the pK, standard ⌬ ؠ ⌬ molar Gibbs energy rG , standard molar enthalpy rH°, and standard molar heat ca- ؠ ⌬ pacity change rC p for each of the ionization reactions at the temperature T ϭ298.15 K and the pressure pϭ0.1 MPa. The standard state is the hypothetical ideal solution of unit molality. The chemical name͑s͒ and CAS registry number, structure, empirical formula, and molecular weight are given for each buffer considered herein. The selection of the values of the thermodynamic quantities for each buffer is discussed. © 2002 by the U.S. Secretary of Commerce on behalf of the United States. All rights reserved. Key words: buffers; enthalpy; equilibrium constant; evaluated data; Gibbs free energy; heat capacity; ionization; pK; thermodynamic properties. Contents 7.19. CAPSO.............................. 263 7.20. Carbonate............................ 264 7.21. CHES............................... 264 1. Introduction................................ 232 7.22. Citrate............................... 265 2. Thermodynamic Background.................. 233 7.23. L-Cysteine............................ 268 3. Presentation of Data......................... 234 7.24. Diethanolamine........................ 270 4. Evaluation of Data. ........................ 235 7.25. Diglycolate.......................... -
Buffers a Guide for the Preparation and Use of Buffers in Biological Systems Calbiochem® Buffers a Guide for the Preparation and Use of Buffers in Biological Systems
Buffers A guide for the preparation and use of buffers in biological systems Calbiochem® Buffers A guide for the preparation and use of buffers in biological systems Chandra Mohan, Ph.D. EMD, San Diego, California © EMD, an affiliate of Merck KGaA, Darmstadt, Germany. All rights reserved. A word to our valued customers We are pleased to present to you the newest edition of Buffers: A Guide for the Preparation and Use of Buffers in Biological Systems. This practical resource has been especially revamped for use by researchers in the biological sciences. This publication is a part of our continuing commitment to provide useful product information and exceptional service to you, our customers. You will find this booklet a highly useful resource, whether you are just beginning your research work or training the newest researchers in your laboratory. Over the past several years, EMD Biosciences has clearly emerged as a world leader in providing highly innovative products for your research needs in Signal Transduction, including the areas of Cancer Biology, Alzheimer’s Disease, Diabetes, Hypertension, Inflammation, and Apoptosis. Please call us today for a free copy of our LATEST Catalog that includes tools for signal transduction and life science research. If you have used our products in the past, we thank you for your support and confidence in our products, and if you are just beginning your research career, please call us and give us the opportunity to demonstrate our exceptional customer and technical service. Corrine Fetherston Sr. Director, Marketing ii Table of Contents: Why does Calbiochem® Biochemicals Publish a Booklet on Buffers? . -
Tris(Hydroxymethyl)Aminomethane; Tris
Tris(hydroxymethyl)aminomethane; Tris Technical Bulletin No. 106B IMPORTANT NOTICE The "Tris" described in this bulletin is Tris(hydroxymethyl)aminomethane. It is not the "Tris" used to flame-proof fabric. The latter, Tris(2,3-dibromopropyl)phosphate, has been reported to be a cancer suspect agent. Trizma® is Sigma's registered trademark applied to various compounds of tris(hydroxymethyl)- aminomethane (tris) prepared by Sigma. For example, Trizma HCl is the completely neutralized crystalline hydrochloride salt of tris. Trizma Base is the pure tris itself. Trizma and its salts have been useful as buffers in a wide variety of biological systems. Uses include pH control in vitro1,2 and in vivo,3,4 for body fluids and as an alkalizing agent in the treatment of acidosis of the blood.5 Trizma has been used as a starting material for polymers, oxazolones (with carboxylic acids) and oxazolidines (with aldehydes).6 Trizma does not precipitate calcium salts and is of value in maintaining solubility of manganese salts.7 PRIMARY BASIMETRIC STANDARD Trizma Base meets many of the requirements for a primary basimetric standard. It is pure, essentially stable, relatively non-hygroscopic and has a high equivalent weight. Trizma Base can be dried at 100°C for up to 4 hours.8 It can be used for the direct standardization of a strong acid solution; the equivalence point can be determined either potentiometrically or by use of a suitable indicator: [3-(4-Dimethylamino-1-naphthylazo)-4- methoxybenzenesulfonic acid, Product No. D5407]. Tris(hydroxymethyl)aminomethane (Trizma Base) -- Physical Data Empirical Formula: C4 H11 N O3 Molecular Weight: 121.14 Equivalent Weight: 121.14 pH of 0.05 M aqueous solution: 10.4* Kb = 1.202 × 10-6 at 25° (pKa = 8.08)9 Trizma HCl -- Physical Data Empirical Formula: C4 H12 Cl N O3 Molecular Weight: 157.6 Equivalent Weight: 157.6 pH of 0.05 M aqueous solution: 4.7* *Trizma HCl in solution will produce a pH of approximately 4.7, but will have little if any buffering capacity. -
Novel Process for the Preparation of Serinol
Europaisches Patentamt 0 348 223 J European Patent Office fin Publication number: A2 Office europeen des brevets EUROPEAN PATENT APPLICATION Application number: 89306393.3 int. a*: C 07 C 89/00 C 07 C 91/12 Date of filing: 23.06.89 Priority: 23.06.88 US 210945 ® Applicant: W.R. Grace & Co.-Conn. 1114 Avenue of the Americas New York New York 1 0036 (US) Date of publication of application: 27.12.89 Bulletin 89/52 @ Inventor: Quirk, Jennifer Maryann 6566 River Clyde Drive Designated Contracting States: 20777 (US) AT BE CH DE ES FR GB GR IT LI LU NL SE Highland Maryland Harsy, Stephen Glen 16236 Compromise Court Mt. Airy Maryland 21771 (US) Hakansson, Christer Lennart Vikingsgaten 3 S-25238Helsingborg (SE) @ Representative: Bentham, Stephen et al J.A. Kemp & Co. 14 South Square Gray's Inn London WC1R5EU (GB) @ Novel process for the preparation of serinol. (g) A process for forming 2-amino-1 ,3-propanediol by reduc- ing a 5-nitro-1,3-dioxane and subsequently hydrolyzing the reduced compound. CO SI 00 3 Q_ LU Bundesdruckerei Berlin EP 0 348 223 A2 Description NOVEL PROCESS FOR THE PREPARATION OF SERINOL The present invention relates to a novel process to form 2-amino-1 ,3-propanediol (commonly known as "serinol"). The present process provides a means of forming serinol using readily formed materials under mild 5 and easily handled conditions suitable for industrial application. Serinol is a highly desired material required for the preparation of nonionic x-ray contrast media, such as iopanediol (N,N'-bis[2-hydroxy-1-(hydroxymethyl)ethyl]-2,4,6-triiodo-5-lactamidisophthalamide). -
Bis-Tris (B4429)
Bis-Tris Cell Culture Tested Product Number B 4429 Product Description Bis-Tris buffer has been utilized in a study of λ cro 7 Molecular Formula: C8H19NO5 repressor self-assembly and dimerization. An Molecular Weight: 209.2 investigation of the ligand binding properties of the CAS Number: 6976-37-0 human hemoglobin variant (Hb Hinsdale) has used 8 pKa: 6.5 (25 °C) Bis-Tris buffer. Synonyms: 2-bis(2-hydroxyethyl)amino-2- (hydroxymethyl)-1,3-propanediol, bis(2- Precautions and Disclaimer hydroxyethyl)amino-tris(hydroxymethyl)methane, For Laboratory Use Only. Not for drug, household or 2,2-bis(hydroxymethyl)-2,2',2''-nitrilotriethanol other uses. This product is cell culture tested (4.2 mg/ml) and is Preparation Instructions designated as Biotechnology Performance Certified. It This product is soluble in water (500 mg/ml), yielding a is tested for endotoxin levels and for the absence of clear, colorless to very faint yellow solution. nucleases and proteases. References Bis-Tris is a zwitterionic buffer that is used in 1. Good, N. E., et al, Hydrogen ion buffers for biochemistry and molecular biology research. It is biological research. Biochemistry, 5(2), 467-477 structurally analogous to the the Good buffers that (1966). were developed to provide buffers in the pH range of 2. Molecular Cloning: A Laboratory Manual, 3rd ed., 6.15 - 8.35 for wide applicability to biochemical Sambrook, J. and Russell, D. W., CSHL Press studies.1 The useful pH range of Bis-Tris is 5.8 - 7.2. (Cold Spring Harbor, NY: 2001), pp. 7.26-7.30. -
Formaldehyde Cross-Linking of Chromatin from Drosophila
2 Formaldehyde Cross-linking of Chromatin from Drosophila Protocol from modEncode IGSB University of Chicago originally written by Alex Crofts and Sasha Ostapenko and updated by Matt Kirkey. 1. Set centrifuge on fast temp to cool down to 4°C (sign out centrifuge). 2. Turn on sonicator, clean probes (sign out sonicator). 3. Prep A1 and Lysis buffers. 4. Collect material, wash with embryo wash buffer. 5. Add material to a Broeck-type homogenizers(Wheaton #357426). Embryo’s can be collected directly in Douncer-type homogenizer(Wheaton #357544). 6. Add 6 mL A1 buffer to Potter homogenizers. 7. Add 500 uL of 20% formaldehyde (to 1.8%) and begin 15’ timer. 8. Homogenize material (100-300mg/IP of embryos, larvae, pupae, adults) first in Broeck homogenizer (frosted) and then in Douncer (clear) with type A pestle (tight) at RT (6 strokes each). 9. Transfer homogenate to 15mL polystyrene tube. Mix by inverting every 5 minutes. 10. Once 15’ timer is up, add 540 uL of 225 mM glycine solution, mix and incubate 5’ at RT. 11. Put on ice and keep on ice unless specified. 12. Centrifuge 5’ at 4000 rpm at 4°C. Discard supernatant. 13. Add 3mL of buffer A1. Resuspend the pellet and centrifuge for 5’ at 4000 rpm and 4°C. Repeat this washing step 3 more times. 14. Wash once in 3 mL of lysis buffer without SDS. Centrifuge for 5’ at 4000 rpm and 4°C. Discard supernatant. 15. Resuspend the cross-linked material in 0.5 mL of lysis buffer. Add SDS to 0.1% (2.5 uL of 20% SDS) and N-lauroylsarcosine to 0.5% (12.5 uL of 20% solution). -
Protein Gel Electrophoresis Technical Handbook Separate Transfer Detect 2
Western blotting Protein gel electrophoresis technical handbook separate transfer detect 2 Comprehensive solutions designed to drive your success Protein gel electrophoresis is a simple way to separate proteins prior to downstream detection or analysis, and is a critical step in most workflows that isolate, identify, and characterize proteins. We offer a complete array of products to support rapid, reliable protein electrophoresis for a variety of applications, whether it is the first or last step in your workflow. Our portfolio of high-quality protein electrophoresis products unites gels, stains, molecular weight markers, running buffers, and blotting products for your experiments. For a complete listing of all available products and more, visit thermofisher.com/separate For orderingordering informationinformation referrefer to pagepage XX.XX ForForqr quickquickrk referencereferencee on the protocolprotocol pleasepleasere referrefertr totopo pagepage XX. Prepare samples Choose the electrophoresis Select precast gel and select buffers Select the standard chamber system and power supply Run the gel Stain the gel Post stain 3 Contents Electrophoresis overview 4 Select precast gel Gel selection guide 8 Gels 10 Prepare samples and select buffers Sample prep kits 26 Buffers and reagents 28 Buffers and reagents selection guide 29 Select the standard Protein ladders 34 Protein standards selection guide 36 Choose the electrophoresis chamber system and power supply Electrophoresis chamber systems 50 Electrophoresis chamber system selection guide 51 -
Nupage Technical Guide
NuPAGE® Technical Guide General information and protocols for using the ® NuPAGE electrophoresis system Rev. date: 29 October 2010 Manual part no. IM-1001 MAN0003188 User Manual 2 Contents ® NuPAGE Precast Gels................................................................................................. 5 General Information.......................................................................................................................................5 Description of the NuPAGE® Electrophoresis System ..............................................................................6 NuPAGE® Gel Specifications ........................................................................................................................8 Gel Selection ....................................................................................................................................................9 Well Volume..................................................................................................................................................10 Gel Staining ...................................................................................................................................................11 Methods ....................................................................................................................... 12 General Guidelines for Samples and Buffers............................................................................................12 Preparing Buffers for Denaturing Electrophoresis ..................................................................................14 -
Guidesheet and Formulas Learly Label All Containers with Content Name
Biological Abbreviations GuideSheet and Formulas learly label all containers with content name. Write out the complete chemical and/or biological name or use abbreviations (useful for small containers). If abbreviations are used, display this guide sheet prominently at C several locations in each lab. Extend the list with lab-specific abbreviations, as necessary. Full Name/Formula Abbreviation Full Name/Formula Abbreviation 2-(N-morpholino)ethanesulfonic acid MES Tris/borate/EDTA TBE 3-(N-morpholino)propanesulfonic acid MOPS Tris/EDTA TE 3-[(3-Cholamidopropyl) Tris/EDTA/sodium chloride TEN CHAPS dimethylammonio]-1-propanesulfonate Tris/glycine/SDS TGS 4-(2-hydroxyethyl)-1- HEPES piperazineethanesulfonic acid Tris/phosphate/EDTA TPE Bovine serum albumin BSA Tris/sodium chloride/Tween 20 TNT Dulbecco's Modified Eagle Medium DMEM Tween 20/TBS TTBS Dulbecco's phosphate-buffered saline DPBS Glycerol, Tris-HCl, EDTA, Bromphenol 6 x DNA Loading dye Blue, Xylene Cyanol FF (DNA Agarose gels) (RT) Ethylene glycol-bis(β-aminoethyl EGTA ether)-N,N,N′,N′-tetraacetic acid Protein A/G A/G (4°) Ethylenediaminetetraacetic acid EDTA Alkaline Phosphatase AP (4°) g,4-Dihydroxy-2-(6-hydroxy-1- Fetal bovine serum FBS BFA – Brefeldin A heptenyl)-4 cyclopentanecrotonic acid Solution (4°) Hanks balanced salt solution HBSS l-lactone HEPES-buffered saline solution HeBS 2-Bis(2-hydroxyethyl)amino-2- Bis-Tris (RT) (hydroxymethyl)-1,3-propanediol piperazine-N,N′-bis(2-ethanesulfonic PIPES acid) Caesium chloride (used for plasmid CsCl2 (RT) prep) Phosphate-buffered -
Buffers for Biochemical Reactions Protocols and Applications Guide
||||||||||||||| 15Buffers for Biochemical Reactions PR CONTENTS I. What a Buffer System Is and How It Works 1 O II. What Makes a "Good" Buffer 1 III. Preparing Buffers 2 T OCOLS A. Prepare Buffers at the Appropriate Temperature and Concentration. 2 B. Adjust the pH of the Buffer System Correctly. 2 C. Take Care of and Use the pH Meter Correctly. 2 D. Random Tips about Buffer Preparation 2 IV. Appendix A: The Henderson-Hasselbalch Equation 3 & V. Appendix B: Composition and Preparation of Common Buffers and Solutions 4 APPLIC A. Preparation of Bicarbonate-Carbonate Buffer (pH 9.2±10.8) 4 B. Preparation of Citrate Buffer (pH 3.0±6.2) 4 C. Preparation of Phosphate Buffer (pH 5.8±8.0 at 25°C) 4 D. Composition of Additional Buffers and Solutions 4 A VI. References 5 TIONS GUIDE Protocols & Applications Guide www.promega.com rev. 12/12 ||||||||||||||| 15Buffers for Biochemical Reactions PR I. What a Buffer System Is and How It Works • Exclusion by biological membranes. This is not important for all biochemical reactions. However, if Buffers often are overlooked and taken for granted by this is an important criterion for your particular laboratory scientists until the day comes when a bizarre O experiment, it is helpful to remember that zwitterionic artifact is observed and its origin is traced to a bad buffer. buffers (positive and negative charges on different T Although mistakes in the composition of buffers have led atoms within the molecule) do not pass through OCOLS occasionally discoveries such as the correct number of biological membranes. -
Buffer Compatibility with Nano DSC
TA Instruments – Technical Note __________________________________________________________________________________ Buffer Compatibility with Nano DSC Colette Quinn, Ph.D. Microcalorimetry, TA Instruments – Waters LLC, 890 West 410 North, Lindon, UT 84042, USA Introduction When using differential scanning calorimetry (DSC), a high quality buffer scan is imperative in order to accurately determine the partial specific heat capacity of a protein sample or when working with low concentrations of biological macromolecules. The reproducibility of the scans in a Nano DSC instrument is a desirable feature and well documented in the application note: High-throughput DSC: A Comparison of the TA Instruments Nano DSC Autosampler SystemTM with the GE Healthcare VP-Capillary DSCTM. One of the key preparatory steps that will ensure the collection of a good background buffer scan is to condition the DSC sample and reference cells with buffer. Proper cell conditioning is accomplished by filling both cells with buffer and then scanning to the upper temperature limit anticipated for scans that will be collected with test samples in this buffer. During the initial conditioning heat scan, the surface of the cells becomes populated with ions from the buffer. These ions physically adsorb to the surface of the cells. The cells will remain conditioned as long as they are either rinsed with water or cleaned with a dilute detergent, such as 1% Contrad®, followed by a water rinse. Because most proteins are removed using these gentle conditions and the conditioning will be left intact, the cell does not need to be pre-conditioned prior to every sample scan. However, if after a sample scan, a particular biological macromolecule can only be cleaned from the sample cell by the thorough cleaning conditions of 4 M sodium hydroxide followed by 50% formic acid, as recommended in the Nano DSC Getting Started Guide, then the cells will need to be reconditioned.