Bio-rad ReadyPrep™ 2-D Starter Kit Manuel d'utilisateur

ReadyPrep™2-D Starter KitInstruction ManualCatalog Number163-2105For technical servicecall your local Bio-Rad office orin the U.S. call 1-800-4BIORAD(

4 Overlay each of the strips with 2 to 3ml of mineral oil to prevent evaporationduring the rehydration process. Addthe mineral oil slowly, by carefull

9Fig. 6 Wetting the electrode wicks with water.Fig. 7 Draining the oil. Fig. 8 Placing the ReadyStrip gel side downin the focusing tray.

Draining the oil essentially washes theoutside surface of the gel, removingunabsorbed protein and resulting inreduced horizontal streaking. If desired

4.4 Completion of IEF.1. When the electrophoresis run has been completed, remove the IPG strips from the focus-ing tray and transfer them gel side up

6 Figure 9 shows the expected pattern when IPG strips of 17 cm, 11 cm, and 7 cm arestained with IEF gel staining solution (catalog # 161-0434) and des

a Remove the kit from the refrigerator and unpack the 2 bottles of equilibration buffer I (WHITE or SILVER caps), the 2 bottles of equilibration buffe

6. Using Table 3, add the indicated volume of complete equilibration buffer II (containingiodoacetamide) to each strip.7. Return the tray to the orbit

3. Remove an IPG strip from the disposable rehydration/equilibration tray and dip briefly intothe graduated cylinder containing the 1X Tris/glycine/SD

5 Using the forceps, carefully push the strip into the well as shown in Figure 13, taking carenot to trap any air bubbles beneath the strip. When push

174.9 STAINING.The 2-D starter kit was designed so that sufficient protein sample loaded in the IEFdimension readily stain second-dimension SDS-PAGE g

Table of ContentsSection 1 Introduction ...1Section 2 Kit Components...

4.10 Imaging.1 To preserve the gel images, the destained gels can be imaged on a densitometer such asBio-Rad’s GS-800 calibrated imaging densitometer

19Fig. 14 C. PROTEAN Ready Gel stained with Bio-Safe stain.

20Figure 15 A. Ready Gel stained with Coomassie Blue R-250 stain.Fig. 15 B. Criterion stained with Coomassie Blue R-250 stain.

21Figure 15 C. PROTEAN Ready Gel stained with Coomassie Blue R-250 stain.

Section 5AppendixA Buffer and Reagent PreparationTo avoid introducing experimental error when using this kit, we suggest using premixed buffersand sta

234. - The first line displays the step number, slope and voltage- Select HRS:MIN (default)- Enter time of 20 min. - Press NEXTS 01 250 VHRS:MIN vhour

2411. - Select METHOD COMPLETEADD inset delete >ENTER STEP # >NEXT >METHOD COMPLETE >12. - The save method screen is displayed when the na

Section 6Product InformationCatalogNumber Product Description2-D Starter Kit accessories163-2105 ReadyPrep 2-D Starter Kit163-2106 ReadyPrep Rehydrati

Bio-Rad Laboratories, Inc.2000 Alfred Nobel Dr., Hercules, CA 94547510-741-10004110009 Rev A

Section 1IntroductionThe ReadyPrep™2-D starter kit was designed as a single-use kit to familiarize first-time users withthe utilization of the Bio-Rad

Overlay Agarose. One bottle containing 50 ml of 0.5% low melting point agarose in 25 mMTris, 192 mM glycine, 0.1% SDS, and a trace of Bromophenol Blue

Section 4Instructions for UseIntroductionThis 2-D starter kit allows you to successfully separate a complex protein extract by 2-D PAGEusing IPG strip

4Rehydration Setup30 minRehydration Setup30 minIPG Rehydration12 hoursIPG Rehydration12 hoursIPG Rehydration12 hoursIsoelectric Focusing5 hoursIsoelec

4.1 Sample Preparation1 Remove the bottle of rehydration/sample buffer, the bottle of nanopure water and the vialof E. coli protein sample from the ki

2. Repeat this process for the remaining samples by pipetting the indicated volume of sample into adjacent channels. It is best to place samples on bo

Gently place the strip gel side down onto the sample as illustrated in Figure 4. The “+”and “pH 4-7” should be legible and positioned at the left side