Product overview Technical/Spec Support/Documents Tips for usage Troubleshooting guidelines Related products Description Q-PAGE™ Bis-Tris Precast Gel is a high-performance and easy to use precast polyacrylamide gel for electrophoresis in Bis-Tris buffer system (MOPS or MES). The optimized gel formula allows Q-PAGE™ Bis-Tris Precast Gel to show improved resolution, accurate results, and an extended shelf-life over conventional Tris-Glycine gels.  Q-PAGE™ Bis-Tris Precast Gels are available in gradient (4 to 12%) and fixed (8% and 12%) concentrations of polyacrylamide in 12-and 15-well formats. Two available cassette sizes, Mini (10 x 8.3 cm) and Midi (10 x 10 cm), are compatible with most popular protein electrophoresis systems. Q-PAGE™ Mini (QP2XXX) Gels are suitable for Bio-Rad® and other systems. Q-PAGE™ Midi (QP3XXX) Gels are suitable for Invitrogen® XCell SureLock® Mini-Cell, Invitrogen® Mini Gel Tank, Hoefer SE260, and other systems. Key FeaturesUser-friendly gel cassette: Numbered and framed wells for sample loading Labeled warning sign and green tape as reminder Enhanced gel performance:  Enhanced band sharpness Better resolution of small proteins  Stable for shipping at ambient temperature Easy compatibility:  Available as homogeneous and adjusted gradient gels for a wide range of protein separation. Compatible with most popular protein electrophoresis systems Storage and stabilityStore Q-PAGE™ Precast Gels at 4°C for periods up to 12 months. Do not freeze Q-PAGE™ Precast Gels. Remove tape and comb before electrophoresis.   TechnicalClear and sharp bands, high resolutionQ-PAGE™ Bis-Tris Precast Gel shows high resolution of protein separation.QP2320 Specifications GelBis-TrisBuffer systemsMOPS and MESFeaturesClear and sharp bands,high resolutionCassette sizeMini Gel(10 X 8.3 cm)Gel dimensions  8.1 x 7.4 x 0.1 cm(W x L x thickness) cmElectrophoresis systemBio-Rad systemsWell format &Capacity15 wells,22 μl/wellGel percentage12 %Accessory trayProduction descriptionTip cardGel removerCassette opener     ManualManual_Q-PAGE™ Bis-Tris Precast Gel, MiniSDSSDS_Q-PAGE™ Precast GelMigration pattern  Setting Up and Running Q-PAGE™ Mini Precast Gel   Removing Q-PAGE Mini Gel from cassette  Setting up gel/membrane sandwich for Western transfer  Recommendations/Tips for Gel Running1. Remove comb and tape before adaption. 2. Use fresh 1X running buffer for the inner cathode chamber. 3. Do not use Tris-Glycine running buffer for Q-PAGE™ Bis-Tris Precast Gels. 4. Rinse the wells before sample loading. Sample Preparation for SDS-PAGE1.      Mix protein sample with 2X sample buffer. 
2.      Heat the diluted samples at 95°C for 5 min or at 70°C for 10 min.3.      Cool the diluted samples to 4°C and spin down the water condensed on tube surface. (If there is high viscosity part at bottom of tube, transfer supernatant to a new tube.) Prepare Q-PAGE™ for Sample Loading1.Open the blister tray of Q-PAGE™ Precast Gel.2.Briefly rinse the gel cassette with ddH2O.3.Remove tape and comb; avoid squeezing the gel.4.Adapt Q-PAGE™ to electrophoresis system; instruction are provided below. (BioRad Mini-PROTEAN® Core Electrophoresis System is recommended.) 5.Use a pipette to gently wash the wells with running buffer to remove residual storage buffer. 6.Fill the wells with running buffer prior to sample loading. 7.Load samples and pre-stained protein marker into numbered wells.8.Fill both inner and outer chambers with running buffer to the highest level. Ensure gel wells are completely covered.  Power Setting for Running Q-PAGE™Optimize the voltage and running time if needed. 130 V180 V230 V*2Running Time*145-60 mins25-40 mins15-30 minsExpected CurrentInitial (per gel)Final (per gel) 60-70 mA20-25 mA 100-110 mA40-50 mA 130-140 mA60-70 mAExpected temperature25-30°C25-35 °C35-45°C*1 Set voltage higher than 100 V is recommended. *2 For higher voltage conditions, please use fresh running buffer for inner and outer chambers. *3 Running time varies depending on gel percentage, running buffer, temperature, and power supplyRemove Q-PAGE™ Gel from CassetteOpen cassette immediately after electrophoresis. Avoid gel drying.1.Insert the cassette opener into corners of cassette. 2.Sequentially pry the opener to separate the two plates. 3.Gently pull two plates apart from the top of cassette.4.Carefully detach the gel either from the bottom of gel or the top side of the cassette.-Avoid diagonally peeling the gel from the corner.       -Use water to help gel detachment if it needed5.Gently remove the gel for further staining or Western blotting.  Gel Staining  Proteins separated using Q-PAGE™ Precast Gels can be further stained with most popular staining reagents, such as Coomassie dyes (R-250 or G-250), Silver-stain solution, and FluoroStain™ Protein Fluorescent Staining Dye. (Cat. No. PS1000)  Transferring Protein from Q-PAGE™ to Blotting Membrane 1. After protein separation using Q-PAGE™, gently detach QPAGE™ from cassette and then equilibrate the gel in transfer buffer. 2. Pre-soak blotting membrane and filter papers in transfer buffer.          3. Assemble transfer sandwich by orientating cathode, sponge, filter papers, gel, membrane, filter papers, sponge, and anode. The protein goes to the direction of cathode to anode. 4. Carefully move roller over the gel/membrane to remove air bubbles and excess buffer until complete contact is established. 5. Insert transfer cassette into transfer module. Notice that black side of cassette should be next to black side of module. 6. Fill transfer tank with pre-cooled transfer buffer to the highest water level. 7. Set constant voltage at 100 V. Transfer for 90 minutes at low temperature condition. Pre-stained protein marker should be visible on the membrane after transfer is completed.     Transfer of proteins to the membrane can be checked using Ponceau S staining before blocking step. Supplemental Information for Using Q-PAGE™ Precast Gel  Adapting Q-PAGE™ Mini Precast Gel to BioRad Mini-PROTEAN® Core 1. After removing comb and tape, place the Q-PAGE™ Mini Precast Gel with notched plate facing toward inner chamber.  2. Align the notched plate to ensure the edge sits just below the notch at the top of green gasket.  3. Gently press gel cassette toward green gasket and then lock gel cassette with two green arms. Avoid squeezing the cassette and gel. 4. Fill inner chamber with running buffer to check tightness of seal. If necessary, reassemble and check the seal again. 5. Fill inner chamber with running buffer to ensure gel wells are completely covered. 6. Fill outer chamber with running buffer to the highest level. Adapting Q-PAGE™ Mini Precast Gels to other electrophoresis system, please follow the manufacturer’s instruction. Buffer recipes 2X sample buffer with reducing agent  62.5 mM Tris-HCl pH 6.8, 2% SDS, 25% (v/v) glycerol, 0.01% bromophenol blue, 5% β-mercaptoethanol or 100 mM DTT (added fresh) 10X MOPS running buffer 60.6 g Tris base, 104.6 g MOPS, 10.0 g SDS, 3.0 g EDTA.  Bring up the volume to 1 L with ddH2O. 10X MES running buffer 60.6 g Tris base, 97.6 g MES, 10.0 g SDS, 3.0 g EDTA.  Bring up the volume to 1 L with ddH2O. 1X running buffer Dilute 100 ml 10X running buffer with 900 ml ddH2O. 10X transfer buffer 30.0 g Tris base, 144.0 g Glycine. Bring up the volume to 1 L with ddH2O. 1X transfer buffer *Cool 1X transfer buffer to 4°C before using. Dilute 100 ml 10X transfer buffer with 200 ml methanol and 700 ml ddH2O. **Add SDS to 0.1% to promote transfer of high molecular weight proteins.    Troubleshooting Guidelines ProblemPossible CauseSuggested Solution Well deformationPull one side of comb out of cassette.Smoothly pull the comb straight out of the cassette. Bubbles between gel and cassetteGel has been frozen or stored at wrong temperature.Store Q-PAGE Precast Gels at 4°C. Buffer leaking from the inner chamberUntight assembly of gels to the electrode modulesReassemble Q-PAGE gels into the electrodemodules.Fill outer chamber with 1X running buffer to thehighest level. Samples do not sink into the wells.Residual gel storage buffer in the wellsRinse the gel wells with ddH2O or 1X running bufferbefore loading. Insufficient sample bufferUse more sample buffer to prepare samples. Current is zero and sample do not migrate into gelTape at bottom of gel not removedRemove tape Gels run faster or more slowly than expected.Incorrect running bufferCheck buffer composition.Use fresh 1X running buffer for inner chamber. Crooked bands at middle or bottom of gelGel has been frozen or stored at wrong temperature.Store Q-PAGE Precast Gels at 4°C. Incorrect running bufferCheck buffer composition.Use fresh 1X running buffer for inner chamber. Band pattern curves toward one or both sides of gel.Buffer leaking from the inner chamberCheck assembly of gels into the electrode modules. Excessive heating of gelCheck buffer composition. Or dilute running bufferto 0.5-0.75X.Do not exceed recommended running conditions. Insufficient buffer in inner or outer buffer chamberFill inner and outer chambers to completely covergel wells. Poor resolution or fuzzy bandsExcessive heating of gelCheck buffer composition.Do not exceed recommended running conditions. Incorrect running bufferCheck buffer composition. Bands are missing on the membrane after Westerntransferring.Proteins move in the wrong directionCheck the order of gel/membrane sandwich assembly,the direction of transfer cassette in transfer modules, and the polarity ofconnections to power supply. Swirls or missing bands; bands trail off in multipledirections on the membrane after Western transferring.Contact between the membrane and the gel was poor;Air bubbles or excess buffer remains between the blotting membrane andthe gel. Use thicker/more filter paper in the gel/membranesandwichRemove air bubbles and excess buffer betweengel and membrane by carefully moving the roller over the membrane. Apparent molecular sizes of prestained proteinmarkers are different as indicated.Prestained protein markers used have not beencalibrated for use with Q-PAGE gels. Dyes for staining protein markers affect themigration patterns of prestained proteins in different buffer systems.Calibrate prestained protein markers againstunstained proteins of known size or use SMOBIO’s ExcelBand™ Protein Markers.  Q-PAGE™ Precast Gel Gel TypeBis-TrisTGN (Tris-Glycine-Novel)Buffer systemsMOPS and MESTris-Glycine (Laemmli)FeaturesClear and sharp bands, high resolutionQuick running, clear bandsCassette sizeMini Gel(10 x 8.3 cm)Midi Gel(10 X 10 cm)Mini Gel(10 x 8.3 cm)Midi Gel(10 X 10 cm)Electrophoresis systemBio-Rad systemsMini Gel TankXcell SureLock,Hoefer SE260Bio-Rad systemsMini Gel TankXcell SureLock,Hoefer SE260Well format &Capacity12 wells,  25 μl/well15 wells,22 μl/well12 wells, 40 μl/well15 wells, 28 μl/well12 wells,  25 μl/well15 wells,  22 μl/well12 wells,  40 μl/well15 wells,  28 μl/wellGel percentage/Cat. No.8%8%8%8%10%10%10%10%QP2110QP2120QP3110QP3120QP4210QP4220QP5210QP522012%12%12%12%4-15%4-15%4-15%4-15%QP2310QP2320QP3310QP3320QP4510QP4520QP5510QP55204-12%4-12%4-12%4-12%    QP2510QP2520QP3510QP3520    ExcelBand™ Protein MarkersReady-to-use— premixed with a loading buffer for direct loading, no need to boilBroad range— 310 kDa to 5 kDaPre-stained bands — for monitoring protein separation during electrophoresis and Western blotting transferring efficiency on membraneEnhanced bands— for quick referenceYesBlot™ Western Marker IReady-to-use — no need of mixing or heating before sample loadingDirect visualization — 10 IgG-binding proteins for direct visualization on Western blotsPre-stained bands — 4 pre-stained proteins for monitoring protein separation during electrophoresis and Western blotting transferring efficiency on membraneWide range — 10 clear bands from 15 to 200 kDa for size estimationQuick reference — two enhanced bands (30 and 80 kDa)FluoroStain™ Protein Fluorescent Staining DyeCompatible to MASS analysis — compatible to the analysis of mass spectra, such as LC-MS/MS, MALDI-TOF, and etc.High sensitivity — detection level achieve ~3 ng, similar to silver stainingSubstitution of the Coomassie Blue protein staining method