ADDENDUM. Ready Gel System RESOURCE GUIDE. Applications:

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1 Ready Gel System RESOURCE GUIDE ADDENDUM This addendum includes new electrophoresis products introduced since the Ready Gel Resource Guide was published and updated information for a few products. Keep this addendum as a quick reference guide, and use it to find the electrophoresis application you re interested in and all related products available from Bio-Rad. Applications: Protein Molecular Weight Analysis Native Protein Analysis Peptide Analysis Isoelectric Point Analysis/IEF Protease Analysis Oligosaccharide Analysis Glycoprotein Analysis dsdna Analysis ssdna, RNA Analysis Blot Processing Buffers

2 NUCLEIC ACIDS: FOR dsdna ANALYSIS Precast Gel Selection Nucleic Acid Sample Buffer, 5x, 10 ml MW Range TBE Standards Resolved (bp) Gel %T 9 Well 10 Well 10 Well/50 µl 12 Well 15 Well Suggested Standard 200-2,000 5% NEW! EZ Load 20 bp Ruler, ,500 10% NEW! EZ Load 100 bp Ruler, ,000 15% AmpliSize DNA Size Standard, 50-2,000 bp 10-2, % DNA Size Standard, bp Ladder 10x TBE Buffer 1 L L NEW! x 1 L Any Nucleic Acid Standards below 2 kb Gel Staining Run conditions per 2 gels: PowerPac 300 (100/120 V) Begin V (C), 26 ma, minutes PowerPac 300 (220/240 V) End V (C), 22 ma Ethidium Bromide Solution, 10 ml, 10 mg/ml Ethidium Bromide Tablets, 10 x 11 mg Silver Stain Plus Kit Silver Stain Kit NUCLEIC ACIDS: ssdna, RNA ANALYSIS Precast Gel and Standard Selection TBE-Urea Sample Buffer, 30 ml MW Range Resolved TBE-Urea Suggested Standards (bases) Gel %T 9 Well 10 Well 12 Well 15 Well 2-D/Prep Standard 50-1,000 5% AmpliSize DNA Size Standard, 50-2,000 bp % DNA Size Standard, bp Ladder % Oligo Standard, mer Any Nucleic Acid Standards below 2 kb 10x TBE Buffer 1 L L NEW! x 1 L Run conditions per 2 gels: PowerPac 300 (100/120 V) Begin V (C), 30 ma PowerPac 300 (220/240 V) End V (C), 20 ma, minutes Gel Staining Ethidium Bromide Solution, 10 ml, 10 mg/ml Ethidium Bromide Tablets, 10 x 11 mg Radiant Red RNA Stain Solution

3 SDS-PAGE: PROTEIN MOLECULAR WEIGHT ANALYSIS Ready Gel and Standards Selection Laemmli Sample Buffer, 30 ml mercaptoethanol, 25 ml MW Range Tris-HCl Suggested SDS-PAGE Standard Resolved Gel %T 9 Well 10 Well 10 Well/50 µl 12 Well 15 Well 2-D/Prep Standard kda 5% High range kda 7.5% High range kda 10% Broad range kda 12% Low range kda 15% Low range kda 18% Low range kda 4-15% Broad range kda 4-20% Broad range kda 8-16% Broad range kda 10-20% Broad range Protein Stain 10x Tris/Glycine/SDS 1 L L NEW! x 1 L PowerPac 300 (100/120 V) PowerPac 300 (220/240 V) Run conditions per 2 gels: Begin V (C), 100 ma End V (C), 60 ma, 35 minutes Bio-Safe Coomassie 1 L NEW! L NEW! Premixed Coomassie R-250 Kit Silver Stain Plus Kit Silver Stain Kit SYPRO Orange Fluorescent Stain Zinc Stain Kit Transfer Equipment, Membranes, and Detection Reagents (see Ready Gel Resource Guide page 54) Trans-Blot/Mini Trans-Blot, Transfer Buffer: 10x Tris/Glycine 1 L L NEW! x 1 L Alternative Detection: Western Blotting Semi-Dry Transfer, Transfer Buffer: 10x Tris/CAPS 1 L NEW! See page 44 of the Ready Gel Resource Guide for more on stains and stain kits

4 NATIVE PAGE: FOR NATIVE PROTEIN ANALYSIS Ready Gel Selection Native Sample Buffer, 30 ml Suggested Tris-HCl SDS-PAGE Standard Gel %T 9 Well 10 Well 10 Well/50 µl 12 Well 15 Well 2-D/Prep Standard 5% Prestained Blue, Broad % Kaleidoscope, Broad % % % % % % % % Use Prestained standards for reference only, not for molecular weight determination. 10x Tris/Glycine Buffer 1 L L NEW! x 1 L PowerPac 300 (100/120 V) PowerPac 300 (220/240 V) Run conditions per 2 gels: Begin V (C), 100 ma End V (C), 60 ma, 35 minutes Protein Stain Bio-Safe Coomassie 1 L NEW! L NEW! Premixed Coomassie R-250 Kit Silver Stain Plus Kit Bio-Rad Silver Stain Kit Tip: Use any stain except Cu and Zn stains for native separations. Cu and Zn stains require SDS in the gel. SYPRO also requires SDS, however, the SYPRO instruction manual includes a protocol for pre-treating native gels with an SDS containing solution prior to staining. Transfer Equipment, Membranes, and Detection Reagents (see Ready Gel Resourse Guide page 55) Trans-Blot/Mini Trans-Blot, Transfer Buffer: 10x Tris/Glycine 1 L L NEW! x 1 L Alternative Detection: Western Blotting Semi-Dry Transfer, Transfer Buffer: 10 x Tris/CAPS 1 L NEW! See page 55 of the Ready Gel Resource Guide for more on transfer equipment

5 SDS-PAGE: FOR PEPTIDE AND SMALL PROTEIN ANALYSIS Ready Gel and Standard Selection Tricine Sample Buffer, 30 ml mercaptoethanol, 25 ml MW Range Tris-Tricine Suggested Standard Resolved Gel %T 9 Well 10 Well 10 Well/50 µl 12 Well 15 Well 2-D/Prep Standard 4-30 kda 16.5% Polypeptide SDS-PAGE 5-40 kda 10-20% Kaleidoscope Polypeptide Standard 10x Tris/Tricine/SDS Buffer 1 L x 1 L PowerPac 300 (100/120 V) PowerPac 300 (220/240 V) Run conditions per 2 gels: Begin V (C), 65 ma End V (C), 35 ma, 100 minutes Protein Stain Bio-Safe Coomassie 1 L NEW! L NEW! Coomassie G-250, 10 g Silver Stain Plus Kit Silver Stain Kit Recommended Silver Stains recommended for peptides larger than 3.5 Kd Transfer Equipment, Membranes, and Detection Reagents (see Ready Gel Resourse Guide page 54) Trans-Blot/Mini Trans-Blot, Transfer Buffer: 10x Tris/Glycine 1 L L NEW! x 1 L Alternative Detection: Western Blotting Semi-Dry Transfer, Transfer Buffer: 10x Tris/CAPS 1 L NEW! See page 38 of the Ready Gel Resource Guide for more on power supplies

6 IEF: ISOELECTRIC POINT ANALYSIS IEF Sample Buffer, 30 ml Ready Gel and Standard Selection IEF Ready ph Range Suggested SDS-PAGE Standard Gels %T Resolved 9 Well 10 Well 10 Well/50 µl 12 Well 15 Well 2-D/Prep Standard 5% ph IEF Standard, pi % ph IEF Standard, pi x Anode Buffer, 250 ml x Cathode Buffer, 250 ml x Formulations: Anode Buffer: 7 mm Phosphoric Acid Cathode Buffer: 20 mm Lysine/20 mm Arginine PowerPac 1000 (100/120 V) PowerPac 1000 (220/240 V) Run conditions per 2 gels stepwise: 100 V (C), 5-25 ma 1 hour, 250 V (C), 5-25 ma 1 hour, 500 V (C), 5-25 ma, 30 minutes Gel Stain Premixed IEF Stain, 1 L Silver Stain Plus Kit Silver Stain Kit Transfer Equipment, Membranes, and Detection Reagents (see Ready Gel Resourse Guide page 54) Alternative Detection: Western Blotting Trans-Blot/Mini Trans-Blot, Transfer Buffer: 0.7% Acetic Acid, see Tip #25, page 55 in the Ready Gel Resource Guide See page 42 of the Ready Gel Resource Guide for more on premixed sample and running buffers

7 ZYMOGRAM: FOR PROTEASE ANALYSIS Ready Gel and Standard Selection Zymogram Sample Buffer, 30 ml MW Range Zymogram Gel %T With Suggested SDS-PAGE Standard Resolved Contents 9 Well 10 Well 10 Well/50 µl 12 Well 15 Well Standard kda 10% with Prestained, low range Gelatin Prestained, broad range kda 12% with Prestained, high range ß-Casein 10x Tris/Glycine/SDS Buffer 1 L L NEW! x 1 L PowerPac 300 (100/120 V) PowerPac 300 (220/240 V) Run conditions per 2 gels: Begin V (C), 25 ma, minutes End V (C), 12 ma Renaturation Zymogram Renaturation Buffer, 10x, 125 ml Development Zymogram Development Buffer, 10x, 125 ml Note: Document gels prior to staining since stain will obscure standards. Gel Staining Coomassie R-250, 10 g (For Zymogram Staining, use as a solution of: 0.5% Coomassie R250, 10% Acetic Acid, 40% methanol) See page 40 of the Ready Gel Resource Guide for more on gel drying equipment

8 OLIGOSACCHARIDE ANALYSIS Step Ready Gel System Product / N-Linked Oligosaccharide Analysis O-Linked Oligosaccharide Analysis Monosaccharide Analysis Glycoprotein in non-tris buffer Glycoprotein in non-tris buffer Glycoprotein in non-tris buffer Kit Selection Profiling Kit / Profiling Kit / Composition Kit / Sequencing Kit / Oligosaccharide Buffer Pack/ Oligosaccharide Buffer Pack/ Monosaccharide Buffer Pack/ Mini-PROTEAN 3 Electrophoresis Mini-PROTEAN 3 Electrophoresis Mini-PROTEAN 3 Electrophoresis Module / Module / Module / Carbohydrate Gaskets Carbohydrate Gaskets Carbohydrate Gaskets PowerPac 300 (100/120 V) PowerPac 300 (100/120 V) PowerPac 300 (100/120 V) PowerPac 300 (220/240 V) PowerPac 300 (220/240 V) PowerPac 300 (220/240 V) Run conditions per 2 gels: Run conditions per 2 gels: Run conditions per 2 gels: Begin: V, 30 ma (C), Begin: V, 40 ma (C), Begin: V, 60 ma (C), minutes minutes minutes End: V, 30 ma (C) End: V, 40 ma (C) End: V, 60 ma (C) ANTS Fluorophore ANTS Fluorophore AMAC Fluorophore Detection Method Imaging Options Gel Doc 2000 or Gel Doc 2000 or Gel Doc 2000 or mini transilluminator mini transilluminator mini transilluminator Visit for information on the new Mini-PROTEAN 3 Electrophoresis System

9 GLYCOPROTEIN ANALYSIS Ready Gel and Standard Selection Laemmli Sample Buffer, 30 ml mercaptoethanol, 25 ml MW Range Tris-HCl Suggested Standard Resolved Gel %T 9 Well 10 Well 10 Well/50 µl 12 Well 15 Well 2-D/Prep Standard kda 5% High range kda 7.5% High range kda 10% Broad range kda 12% Low range kda 15% Low range kda 18% Low range kda 4-15% Broad range kda 4-20% Broad range kda 8-16% Broad range kda 10-20% Broad range x Tris-Glycine/SDS Buffer 1 L x Tris-Glycine/SDS Buffer 5 L NEW! PowerPac 300 (100/120 V) PowerPac 300 (220/240 V) Run conditions per 2 gels: Begin V (C), 100 ma End V (C), 60 ma, 35 minutes Glycoprotein Detection Colorimetric Western Blot (BCIP/NBT) Detection Method Deglycosylation Detection Coomassie R-250 Staining Solutions Kit See page 38 of the Ready Gel Resource Guide for more on power supplies

10 BLOT PROCESSING BUFFERS Sample Method of Transfer Buffer Stock Volume Proteins Trans-Blot/Mini Trans-Blot 10x Tris/Glycine 1 L L NEW! x 1 L Proteins Trans-Blot Semi-Dry 10x Tris/CAPS (requires SDS) 1 L ml DNA/RNA Trans-Blot/Mini Trans-Blot 10x TBE 1 L L NEW! x 1 L x TAE 1 L L NEW! x 1 L DNA/RNA Trans-Blot Semi-Dry 10x TBE 1 L L NEW! x 1 L DNA/RNA Capillary transfer 20x SSC 1 L NEW! L NEW! Item Product Package Size Wash buffers 10x TBS 1 L NEW! x PBS 1 L NEW! x SSC 1 L NEW! L NEW! Blocking Buffers 1x TBS/1% Casein 1 L NEW! x PBS/1% Casein 1 L NEW! % Tween 1 L NEW! Ready Gels Accelerate Your Research! Bio-Rad Laboratories Life Science Group Website Bio-Rad Laboratories Main Office 2000 Alfred Nobel Drive, Hercules, CA 94547, Ph. (510) , Fx. (510) Also in: Australia Ph , Fx Austria Ph. (01) , Fx. (01) Belgium Ph , Fx Canada Ph. (905) , Fx. (905) China Ph /51, Fx Denmark Ph , Fx Finland Ph. 358 (0) , Fx. 358 (0) France Ph , Fx Germany Ph , Fx Hong Kong Ph , Fx India Ph. (91-11) , Fx. (91-11) Israel Ph , Fx Italy Ph , Fx Japan Ph , Fx Korea Ph , Fx Latin America Ph , Fx Mexico Ph , Fx The Netherlands Ph , Fx New Zealand Ph , Fx Russia Ph , Fx Singapore Ph , Fx Spain Ph , Fx Sweden Ph. 46 (0) , Fx. 46 (0) Switzerland Ph , Fx United Kingdom Ph , Fx Bulletin 2364 US/EG Rev A Sig