5 rammentazione dei Peptidi x n-i v n-i y n-i z n-i y n-i-1 -HN-CH-CO-NH-CH-CO-NH- R i a i b i c i wn-i CH-R R i+1 i+1 b i+1 low energy fragmentation d i+1 high energy fragmentation
6 rammentazione dei Peptidi x n-i y n-i z n-i y n-i-1 -HN-CH-CO-NH-CH-CO-NH- R i a i b i c i CH-R R i+1 i+1 b i+1
8 rammentazione dei Peptidi Informazioni complementari x n-i b 1 -a 1 = CO (28) c 1 -b 1 = NH (15) y 1 -x 1 = CO (28) z 1 -y 1 = NH (15) y 2 -y 1 =NHCH(CHR R )CO b 2 -b 1 = NHCH(CHR R )CO -HN-CH-CO-NH-CH-CO-NH- a i R i b i c i y n-i y z n-i-1 n-i CH-R R i+1 i+1 b i+1
9 S/MS validation he large correct number assignment of computational of such a approaches spectrum to and a peptide software tools have equence en developed a first to and automatically central step assign in proteomic peptide sequences data processing. to fragment n spectra. These can be classified in three categories: Database searching, where peptide sequences are identified by correlating acquired fragment ion spectra with theoretical spectra predicted for each peptide contained in a protein sequences database. De novo sequencing, where peptide sequences are explicitly read out directly from fragment ion spectra. Hybrid approaches, such as those based on the extraction of short sequence tags of 3-5 residues in length, followed by error tolerant database searching.
10 Sequenziamento manuale la sequenza aminoacidica viene dedotta irettamente dagli spettri MS o MS/MS. Sequenziamento manuale??? Ma siamo matti? E allora a che serve il computer?!?
11 MS/MS Frammentazione dei Peptidi Individuare gli ioni di sequenza della β-endorfina (MW= )
12 Amino acids Glicine Alanine Serine Proline Valine Threonine Cysteine Isoleucine Leucine Asparagine Aspartic acid Glutamine Lesine Glutamic acid Methionine Histidine Phenylalanine Arginine Tyrosine Tryptophan Gly Ala Ser Pro Val Thr Cys Ile Leu Asn Asp Gln Lys Glu Met His Phe Arg Tyr Trp G A S P V T C I L N D Q K E M H F R Y W Residue mass Immonium ion mass (-) (-) (+) (++) (++) (+) (-) (++) (++) (+) (+) (+) ( ) (+) (+) (++) (++) (-) (++) (+)
13 [M+H] + = m= = Leucine Leu L (++) fragment: b 17
14 [M+H] + = m= = Threonine Thr T (+) fragment: b 16
15 [M+H] + = m = =99.07 Valine Val V (++) fragment: b 15
16 [M+H] + = m = = Leucine Leu L (++) Proline Pro P (++) fragment: b 13
17 [M+H] + = m = = Threonine Thr T (+) fragment: b 12
18 [M+H] + = m = = Glutamine Gln Q (+) fragment: b 11
19 [M+H] + = m = = Serine Ser S (+) fragment: b 10
20 S/MS validation ndividuare gli ioni di sequenza del peptide RPKPQQFFGLM Spettro teorico
21 Amino acids Glicine Proline Leucine Glutamine Lesine Methionine Phenylalanine Arginine Gly Pro Leu Gln Lys Met Phe Arg G P L Q K M F R Residue mass Immonium ion mass (-) (++) (++) (+) ( ) (+) (++) (-) Y 11 Y 10 Y 9 Y 8 Y 7 Y 6 Y 5 Y 4 Y 3 Y 2 Y R P K P Q Q F F G L M b 1 b 2 b 3 b 4 b 5 b 6 b 7 b 8 b 8 b 9 b 10
22 Y 11 Y 10 Y 9 Y 8 Y 7 Y 6 Y 5 Y 4 Y 3 Y 2 Y P K P Q Q F F G L M b 1 b 2 b 3 b 4 b 5 b 6 b 7 b 8 b 9 b 10 b 11
23 OP-DOWN APPROACH: MS/MS validation SAMPLE Remove protein from gel spot 2D PAGE MALDI of peptides mixture Enzymatically digest all protein from spot Search peptide mass MALDI MS/MS Protein MS/MS Ion Fragmentation pattern MS/MS DATABASE SEARCHING
24 ROTEOMIC APPROACHES 1. Top down approach 2. Bottom up or shotgun approach Top down approach: a protein is separated from a complex mixture, purified and identified by direct fragmentation by mass spectrometry Bottom up approach: a mixture of protein is first made more complex through enzymatic digestion, e.g. trypsin, followed by liquid chromatography tandem MS to identify the peptide fragments.
25 OP-DOWN Protein Mixture Run gel; stain; scan Excise spot; wash; digest Simple peptides mixtures 2D SDS PAGE xtract peptides mass analyze Database search
26 OTTOM-UP Protein Mixture Proteolysis Multidimensional LC MS and MS/MS Complex peptides mixture MS/MS Database search
27 dvanced Sample reparation for Proteomics approaches for MALDI
28 Sample Dilution/Concentration ilute samples to the concentrations shown in the table below f the sample concentration is unknown a dilution series may e needed to produce a good spot on the MALDI plate. Compound Peptides and proteins Concentration 0.1 to 10 pmol/µl Oligonucleotides Polymers 10 to 100 pmol/µl 100 pmol/µl Note: highly dilute samples can be concentrated by Speed-Vacuum or Solid Phase Extraction.
29 Sample clean-up Removal of buffer salts, urea, guanidine, EDTA, glycerol, DMSO, detergents, etc. Dilution Washing Drop dialysis Cation exchange Pipette tip column chromatography ZipTips
30 Sample Dilution Simplest way to minimize effect by contaminants. Goal is to dilute contaminants to the point where they no longer interfere with analysis of sample. Requires high enough analyte concentration in sample to provide acceptable data when diluted out.
32 On-Plate Washing Buffer and Salt Removal Dry sample and matrix Deposit 1-2 ml cold 0.1% TFA Leave on for 5-10 sec., then remove Detergent contamination Use 5% Isopropanol Cell Extract Contamination Use 100% Isopropanol
33 Drop Dialysis To remove low molecular weight contaminants Use Millipore membrane, type VS, pore size µm, diam. 25 mm Fill a ml container with deionized water. Float the membrane on the water (shiny side up). Place about 10 ml of sample solution on the membrane. Add 1mL ACN to the sample spot to increase surface area. Allow to sit for ~45 minutes. Remove an aliquot with pipette, add matrix and spot plate.
34 Drop dialysis cleanup of Enolase Before Yeast Enolase (47 kda) in 8 M urea was dialyzed for 1 hr on a Millipore membrane. After
35 Sample Cleanup by Solid Phase Extraction ZipTip -miniature C 18 column chromatography 1. Sample Concentration and Buffer Removal 2. Fractionation 3. Affinity Experiments
36 ssorbimento su Reversed Phase (C18) di peptidi generati da in gel digestion ZipTip: puntali per pipette eppendorf alla cui punta è impaccata una resina che supporta un fase inversa (tipo C18) sulla quale, dopo che i peptidi eluiti dall in.gel digestion (sciolti i ambiente acquoso), gli stessi peptidi si legano. E possibile, con i peptidi legati alla resin effettuare una serie di lavaggi in modo da eliminare eventuali contaminanti ed eluir successivamente i peptidi. (Si adottano le stesse strategie di eluizioni che si adottano i cromatografia HPLC a fase inversa. Questa procedura viene utilizzata soprattutto quando hanno piccoli volumi e si vuole eliminare la presenza di sali.
37 Procedure for using C 18 ZipTips Condition the ZipTip with 10 µl of acetonitrile (ACN), then 10 µl of 50% ACN/0.1% TFA, then 2 x 10 µl of 0.1% TFA. Load the sample onto the ZipTip by pipetting 5-10 µl sample up and down several times and discarding the liquid. Wash C 18 tip with 3 x 10 µl of 0.1% TFA to remove salts. Elute the sample from the ZipTip with 30-70% ACN or elute directly into the matrix (e.g. CHCA in 50% ACN/0.1%TFA); minimal volume of ~3 µl can be used.
38 Use of the C 18 ZipTip 1. Sample Concentration and Buffer Removal 2. Fractionation As peptides and proteins have differing affinities for C 18, the C 18 tips can be used to fractionate mixtures according to their hydrophobicities. Increasing the ACN in a step gradient of 10% - 50% in the eluent increased the number of peptides seen. By fractionating a peptide mass map this can also be beneficial for PSD analysis. 3. Affinity Experiments
39 Step elution with Increasing ACN of IgG HC Endo Lys C Digest from C 18 tip % ACN Counts % ACN % ACN Mass (m/z) Most peptides are seen at 30 % which is a good concentration to use for most digests as this can be used to remove Coomassie Blue which elutes at 40%.
40 In-Gel Digest Fundamentals Handling the gel and slices Washing and destaining Enzymatic Digestion Peptide Extraction Concentration/Cleanup MALDI-TOF Analysis
41 In-Gel Digest Method Success depends upon: Avoiding contamination of samples Digesting the protein efficiently Maximizing recovery of peptides Minimizing losses from handling
42 In-Gel Digest Method Handling the Gel and Slices Gloves and lab coats must be worn at all times to avoid keratin contamination. Work on a clean surface. Use clean polypropylene microcentrifuge tubes, 500 or 1500 ul with snap caps. Test first to confirm OK (i.e., does not leach out polymers, mold release agents, plasticizers, etc.) Set aside a box for digest use only, handle only with gloves. Use only clean tools, containers and reagents for anything that will come in contact with the samples. Keep samples capped at all times unless being processed.
43 In-Gel Digest Method Silver Stained Gels Non-destructive Silver stained samples should be destained prior to analysis as follows: Prepare stock solutions of 30 mm Potassium Ferricyanide and 100 mm Sodium Thiosulfate. Store each at 4 C for up to 3 months. Make the working destain solution immediately prior to use by mixing the two stock solutions above at a 1:1 ratio. Soak gel slices in 100 ul destain solution for 10 minutes. This step converts the silver to a water soluble form. The gel will clear. Carefully remove the destain solution and wash 3X in dh 2 0 (400 ul, 15 min.) Use gel loading tips to prevent accidental aspiration of gel pieces. This step washes away the soluble silver. Ref: Electrophoresis 1999, 20,
44 In-Gel Digest Method Washing Destained Silver and Coomassie Gels Trim the gel slices as needed to approx 1 mm 3. Run a negative and positive control, as well as a reagent control (containing no gel slice). Transfer gels to 500 or 1500 ul capped microcentrifuge tubes Wash gels 3X in 50% ACN / 25 mm NH 4 Bicarbonate ph 8.0 (400 ul, 15 min. each time). This will remove excess Coomassie Blue. Soak in 100% ACN for 5 min. to dehydrate the gels, they will turn opaque white. Remove the ACN. (Note: Be sure that the ACN used does not contain any acid, otherwise the ph will be incorrect. Dry gels in Speed-Vac for min. This will shrink the gels. (Be sure that the inside of the Speed-Vac is clean and free of particulates. Do not allow anyone to use the Speed-Vac with ungloved hands during this step as sample tubes will be uncapped).
45 In-Gel Digest Method Enzymatic Digestion Trypsin Promega Sequencing Grade Modified Trypsin ug/ml in 25 mm NH 4 Bicarbonate ph 8.0. Store at -70 C in onetime-use aliquots. (100 ul each) Rehydrate the dried gels with approx ul cold Trypsin solution. The gels will swell and turn clear. Check after 30 min. for sufficient volume to completely wet entire gel. Add additional Trypsin if needed for large gel pieces. There is no need to overlay with additional buffer. Incubate tightly capped at 37 C for hours. Convection oven is preferable to heat block.
46 In-Gel Digest Method Extraction of Peptides Soak the gel slice in ul 50% ACN / 5% TFA for min. with gentle agitation. Do not vortex. Transfer the supernatant to a second clean tube. Extract the gel again with another ul aliquot of 50% ACN/5% TFA for min. Combine the two extracts and Speed-Vac to complete dryness, about 1 hour. Note: dry at room temp or heat to no more than 30 C. Drying can also be done in a lyophilizer. (Note: Peptides can alternatively be extracted from the gels with % TFA alone if ACN is undesirable, e.g. if ZipTip cleanup will be used.)
48 Staining Procedure Results have shown that Coomassie Blue should be used if the sensitivity is adequate as the recovery of peptides is better than with Silver Staining. Excising the Gel Spot Care should be taken to cut precisely around the stained area to prevent any unnecessary contamination.
BC334 (Proteomics) Practical 1 Calculating the charge of proteins Aliphatic amino acids (VAGLIP) N H 2 H Glycine, Gly, G no charge Hydrophobicity = 0.67 MW 57Da pk a CH = 2.35 pk a NH 2 = 9.6 pi=5.97 CH
Pipe Cleaner Proteins GPS: SB1 Students will analyze the nature of the relationships between structures and functions in living cells. Essential question: How does the structure of proteins relate to their
p H 4 Titration Curve of an Amino Acid Simple amino acid Acidic amino acid Basic amino acid 7 OH - equivalents Objectives: A) To determine the titration curve for an amino acid and B) to use this curve
IV. -Amino Acids: carboxyl and amino groups bonded to -Carbon A. Acid/Base properties 1. carboxyl group is proton donor! weak acid 2. amino group is proton acceptor! weak base 3. At physiological ph: H
Amino Acids, Peptides, Proteins Functions of proteins: Enzymes Transport and Storage Motion, muscle contraction Hormones Mechanical support Immune protection (Antibodies) Generate and transmit nerve impulses
Advanced Medicinal & Pharmaceutical Chemistry CHEM 5412 Dept. of Chemistry, TAMUK Dai Lu, Ph.D. firstname.lastname@example.org Tel: 361-221-0745 Office: RCOP, Room 307 Drug Discovery and Development Drug Molecules Medicinal
WORKING WITH PEPTIDES 1 Synthetic custom peptides offer an increasingly affordable approach for exploring protein-protein interactions and more complex phenomena such as immune responses directed against
HIS-Select Nickel Affinity Gel Catalog Number P6611 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description HIS-Select Nickel Affinity Gel is an immobilized metalion affinity chromatography (IMAC)
Essential rganic Chemistry Chapter 16 The rganic Chemistry of Amino Acids, Peptides, and Proteins Amino Acids a-amino carboxylic acids. The building blocks from which proteins are made. H 2 N C 2 H Note:
PROTEIN SEQUENCING First Sequence The first protein sequencing was achieved by Frederic Sanger in 1953. He determined the amino acid sequence of bovine insulin Sanger was awarded the Nobel Prize in 1958
ProteoSilver Plus Silver Stain Kit Product Code PROT-SIL2 Store at Room Temperature TECHNICAL BULLETIN Product Description The ProteoSilver Plus Silver Stain Kit is an exceptionally sensitive, MALDI-MS
CHEMISTRY ATAR COURSE DATA BOOKLET 2016 Copyright School Curriculum and Standards Authority, 2016 This document apart from any third party copyright material contained in it may be freely copied, or communicated
Guide to Reverse Phase SpinColumns Chromatography for Sample Prep www.harvardapparatus.com Contents Introduction...2-3 Modes of Separation...4-6 Spin Column Efficiency...7-8 Fast Protein Analysis...9 Specifications...10
Guidelines for Writing a Scientific Paper Writing an effective scientific paper is not easy. A good rule of thumb is to write as if your paper will be read by a person who knows about the field in general
Amino Acids & Proteins Shu-Ping Lin, Ph.D. Institute te of Biomedical Engineering ing E-mail: email@example.com Website: http://web.nchu.edu.tw/pweb/users/splin/ edu tw/pweb/users/splin/ Date: 10.13.2010
Application Note Determination of Amino acids by UHPLC with automated PA- Derivatization by the Autosampler Category Bio Analysis Matrix - Method UHPLC Keywords Proteinogenic Amino acids, Canonical Amino
ChemActivity 46 Amino Acids, Polypeptides and Proteins 1 ChemActivity 46 Part A: Amino Acids and Peptides (Is the peptide IAG the same as the peptide GAI?) Model 1: The 20 Amino Acids at Biological p See
BUFFERS and MEDIAS Coomassie Blue Staining Solution 2 g Coomassie Blue 2 L Methanol or Ethanol * 1.6 L 400 ml Glacial acetic acid *If you will be microwaving the gel in staining solution for rapid staining
Mass Standards Kit for Calibration of AB SCIEX TOF/TOF Instruments Protocol 1 Product Description The Mass Standards Kit includes reagents needed to test instrument function, optimize instrument parameters,
Questions- Proteins & Enzymes A. A peptide with 12 amino acids has the following amino acid composition: 2 Met, 1 Tyr, 1 Trp, 2 Glu, 1 Lys, 1 Arg, 1 Thr, 1 Asn, 1 Ile, 1 Cys Reaction of the intact peptide
Human Tubal Fluid (HTF) Media & Modifi ed Human Tubal Fluid (mhtf) Medium with Gentamicin HTF Media are intended for use in assisted reproductive procedures which include gamete and embryo manipulation
292PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 firstname.lastname@example.org A Geno Technology, Inc. (USA) brand name Classic Immunoprecipitation Utilizes Protein A/G Agarose for Antibody Binding (Cat.
Protein Precipitation Protocols Notes: All reagents need to high purity/hplc quality. All tubes used should be new or hand cleaned thoroughly with Micro90 detergent. High quality water needs to be used
Rapid and Reproducible Amino Acid Analysis of Physiological Fluids for Clinical Research Using LC/MS/MS with the atraq Kit Fast, simple and cost effective analysis Many areas of biochemical research and
Protocol SpikeTides TM Peptides for relative and absolute quantification in SRM and MRM Assays Contact us: InfoLine: +49-30-6392-7878 Order per fax: +49-30-6392-7888 or e-mail: www: email@example.com www.jpt.com
June 2010 OXYTOCIN : ADOPTED TEXT FOR THE INTERNATIONAL PHARMACOPOEIA Final text for addition to The International Pharmacopoeia (June 2010) This monograph was adopted at the Forty-fourth WHO Expert Committee
Supporting Information Minimum active structure of insulin-like peptide 5 (INSL5) Alessia Belgi 1,2, Ross A.D. Bathgate *1,2,3, Martina Kocan *4, Nitin Patil 1,5, Suode Zhang 1, Geoffrey W. Tregear 1,2,
Chromatin Immunoprecipitation (ChIP) Day 1 A) DNA shearing 1. Samples Dissect tissue (One Mouse OBs) of interest and transfer to an eppendorf containing 0.5 ml of dissecting media (on ice) or PBS but without
John E. McMurry www.cengage.com/chemistry/mcmurry Chapter 26 Biomolecules: Amino Acids, Peptides, and Proteins Proteins Amides from Amino Acids Amino acids contain a basic amino group and an acidic carboxyl
Online Data Supplement Materials and Methods The ECLIPSE Cohort Patients with COPD were recruited according to the inclusion criteria; aged 40-75, current or ex-smokers with >10 pack-year history; a post
Western Blotting: Mini-gels Materials a Protein Extraction Buffer (for callus or kernel), Solution Stock Final Volume Tris-HCl ph 80 1 M 200 mm 20 ml NaCl 4 M 100 mm 25 ml Sucrose 2 M 400 mm 20 ml EDTA
EXPERIMENT VI PURIFICATION AND CHARACTERIZATION OF PROTEINS I- Protein isolation and dialysis In order to investigate its structure and properties a protein must be obtained in pure form. Since proteins
Problem 1. (12 points total, 4 points each) The molecular weight of an unspecified protein, at physiological conditions, is 70,000 Dalton, as determined by sedimentation equilibrium measurements and by
SPARK DNA Sample Prep Kit Ion Torrent (SPK0002-V08) Frequently Asked Questions Under what circumstances would I use SPARK DNA Sample Prep Kit for Ion Torrent? Enzymatics SPARK DNA Sample Prep Kit for Ion
CONFIRMATION OF ZOLPIDEM BY LIQUID CHROMATOGRAPHY MASS SPECTROMETRY 9.1 POLICY This test method may be used to confirm the presence of zolpidem (ZOL), with diazepam-d 5 (DZP-d 5 ) internal standard, in
Biochemistry - I Prof. S. Dasgupta Department of Chemistry Indian Institute of Technology, Kharagpur Lecture-11 Enzyme Mechanisms II In the last class we studied the enzyme mechanisms of ribonuclease A
TE EMIAL SYTESIS F PEPTIDES Peptides are the long molecular chains that make up proteins. Synthetic peptides are used either as drugs (as they are biologically active) or in the diagnosis of disease. Peptides
Reiner Westermeier, Torn Naven Hans-Rudolf Höpker Proteomics in Practice A Guide to Successful Experimental Design 2008 Wiley-VCH Verlag- Weinheim 978-3-527-31941-1 Preface Foreword XI XIII Abbreviations,
Chromatin Immunoprecipitation A) Prepare a yeast culture (see the Galactose Induction Protocol for details). 1) Start a small culture (e.g. 2 ml) in YEPD or selective media from a single colony. 2) Spin
Studier media 1 Studier Method for Induction Date: 02/09/05, 6/15/05, 8/10/05 Author: based on Bill Studier s Protocol Edited by: R. Kim Revised by: B.Martinez Materials/Reagents/Equipment Reagents Vector:
EXPERIMENT IX Marmara Üniversitesi DETERMINATION OF N-TERMINAL AMINO ACID RESIDUE OF PROTEINS BY THIN LAYER CHROMATOGRAPHY Functions of the proteins depend upon its amino acid sequence. Because amino acid
High-Throughput 3-D Chromatography Through Ion Exchange SPE Application Note 205 Luke Roenneburg and Alan Hamstra (Gilson, Inc.) Introduction 2-dimensional (2-D) separation is the separation of a sample
Running protein gels and detection of proteins 1. Protein concentration determination using the BIO RAD reagent This assay uses a colour change reaction to give a direct measurement of protein concentration.
Application Update Now sold under the Thermo Scientific brand Separation of Peptides from Enzymatic Digestion on Different claim Columns: A Comparative Study INTRODUCTION Separation of peptides which can
User Guide for Reversed-Phase Pipette Tips PACKARD BIOSCIENCE MultiPROBE II COMPATIBLE Notice The information in this document is subject to change without notice and should not be construed as a commitment
Training on the ZQ Open access MS Questions? Contact Dr. Allis Chien firstname.lastname@example.org 650-723-0710 0710 STANFORD UNIVERSITY MASS SPECTROMETRY STANFORD UNIVERSITY MASS SPECTROMETRY 333 CAMPUS DR., MUDD
Proteomics Workflows & Technologies Proteomics "The analysis of the entire PROTEin complement expressed by a genome, or by a cell or tissue type." Wasinger VC et al, Electrophoresis 16 (1995) Proteomics
A Guide to the Analysis and Purification of Proteins and Peptides by Reversed-Phase HPLC HPLC Columns David Carr Your decision has lasting effects. Choose wisely. HPLC Columns Ultra Inert Base-Deactivated
Protein Purification and Analysis Numbers of genes: Humans ~40,000 genes Yeast ~6000 genes Bacteria ~3000 genes Solubility of proteins important for purification: 60-80% soluble, 20-40% membrane Some proteins
Southern Blot Analysis (from Baker lab, university of Florida) DNA Prep Prepare DNA via your favorite method. You may find a protocol under Mini Yeast Genomic Prep. Restriction Digest 1.Digest DNA with
WESTERN BLOTTING TIPS AND TROUBLESHOOTING GUIDE TIPS FOR SUCCESSFUL WESTERB BLOTS TROUBLESHOOTING GUIDE 1. Suboptimal protein transfer. This is the most common complaint with western blotting and could
PowerFecal DNA Isolation Kit Catalog No. Quantity 12830-50 50 Preps Instruction Manual Inhibitor Removal Technology (IRT) is a registered trademark of MO BIO Laboratories, Inc. and is covered by the following
Western Blotting Sive Lab Protocol March 2007 Prepare samples: For zebrafish embryos: Option 1: Take live embryos and put into 1.5 ml tube with E3. Centrifuge gently for 1-2 minutes -yolk lipids will rise
Method Development of LC-MS/MS Analysis of Aminoglycoside Drugs: Challenges and Solutions authors Angela (Qi) Shen, Ling Morgan, Marcele L. Barroso, and Xin Zhang; Tandem Labs Tuyen Nguyen; Sepracor Inc.
Page 1 of 7 Agencourt RNAdvance Cell v2 Total RNA Isolation from Cultured Cells Please refer to http://www.agencourt.com/technical for updated protocols and refer to MSDS instructions http://www.beckmancoulter.com/customersupport/msds/msds.asp
FluoroTag FITC Conjugation Kit Product Number FITC1 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description The FluoroTag FITC Conjugation Kit is suitable for the conjugation of polyclonal and
TRI Reagent For processing tissues, cells cultured in monolayer or cell pellets Catalog Number T9424 Store at room temperature. TECHNICAL BULLETIN Product Description TRI Reagent is a quick and convenient
Thermo Scientific Pierce Controls and Standards for Mass Spectrometry calibration tools for mass spectrometry Ensure confidence in instrument performance with Thermo Scientific Pierce Calibration Solutions
chromatography Thermo Scientific HyperSep Solid Phase Extraction Method Development Guide The following guide provides considerations, tips and general guidelines for developing SPE methods using the Thermo
ProteinChip Energy Absorbing Molecules (EAM) Instruction Manual Catalog #C30-00001, #C30-00002, #C30-00003, #C30-00004 For technical support, call your local Bio-Rad office, or in the US, call 1-800-4BIORAD
Protocol for Western Blotting Materials Materials used on Day 3 Protease inhibitor stock: 1 μg/μl pepstatin A in DMSO 200 μm leupeptin in OG Buffer 200 mm PMSF: Freshly made. Ex) 34.8 mg PMSF in 1 ml isopropanol
PHOENIX PHARMACEUTICALS, INC. The Peptide Elite ASSAY PROTOCOL for INSL3 (Human) RIA Kit (Range:1-128pg/tube) TABLE OF CONTENTS Introduction Contents Storage General Information Assay Conditions General
APPLICATION NOTE POROS CaptureSelect affinity chromatography columns POROS CaptureSelect affinity columns for highspeed quantification of IgG Fc fusion proteins Introduction POROS columns, containing highperformance
Plant Genomic DNA Extraction using CTAB Introduction The search for a more efficient means of extracting DNA of both higher quality and yield has lead to the development of a variety of protocols, however
Methods for Protein Analysis 1. Protein Separation Methods The following is a quick review of some common methods used for protein separation: SDS-PAGE (SDS-polyacrylamide gel electrophoresis) separates
Monoclonal ntibody Fragment Separation and haracterization Using Size Exclusion hromatography oupled with Mass Spectrometry uthors Haiying hen Katherine McLaughlin Sepax Technologies, Inc. 5 Innovation
ABRF 2004 Poster# P55-S Displacement Chromatography Effects Can Cause Highly Selective Sampling of Peptides During Solid Phase Extraction Cleanup by Andrew J. Alpert PolyLC Inc./ 9151 Rumsey Road, ste.
Professor Stuart Conway Introduction to Chemical Biology University of xford Introduction to Chemical Biology ecommended books: Professor Stuart Conway Department of Chemistry, Chemistry esearch Laboratory,
User Protocol 539551 Rev. 16-September-04 JSW Page 1 of Protein Kinase Assay Kit, Universal Cat. No. 539551 Introduction Protein kinases catalyze the transfer of gamma phosphate from adenosine triphosphate
Introduction to Proteomics Why Proteomics? Same Genome Different Proteome Black Swallowtail - larvae and butterfly Biological Complexity Yeast - a simple proteome 6,113 proteins = 344,855 tryptic peptides
AMINO ACIDS & PEPTIDE BONDS STRUCTURE, CLASSIFICATION & METABOLISM OBJECTIVES At the end of this session the student should be able to, recognize the structures of the protein amino acid and state their
Electrospray Ion Source Electrospray Ion Trap Mass Spectrometry Introduction The key to using MS for solutions is the ability to transfer your analytes into the vacuum of the mass spectrometer as ionic
Protein physics, Lecture 5 Peptide bonds: resonance structure Properties of proteins: Peptide bonds and side chains Proteins are linear polymers However, the peptide binds and side chains restrict conformational