![SMOBIO/[QP3120] Q-PAGE™ Bis-Tris Precast Gel (Midi, 15 wells, 8%), 10 gels/Midi, 15 wells, 8%), 10 gels</span>
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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 Features
User-friendly gel cassette:
Numbered and framed wells for sample loading
With cassette opener for easy use
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 stability
Store 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.
Clear and sharp bands, high resolution
Q-PAGE™ Bis-Tris Precast Gel shows high resolution of protein separation.
QP3120 Specifications
Gel | Bis-Tris | |
Buffer systems | MOPS and MES | |
Features | Clear and sharp bands, high resolution | |
Cassette size | Midi Gel (10 X 10 cm) | |
Gel dimensions | 8.1 x 8.1 x 0.1 cm (W x L x thickness) cm | |
Electrophoresis system | Mini Gel Tank XCell SureLock, Hoefer SE260 | |
Well format & Capacity | 15 wells, 28 μl/well | |
Gel percentage | 8 % | |
Accessory tray | Production description Tip card Gel remover Cassette opener | |
Manual
Manual_Q-PAGE™ Bis-Tris Precast Gel, Midi
SDS
SDS_Q-PAGE™ Precast Gel
Migration pattern
Setting Up and Running Q-PAGE™ Midi Precast Gel
Removing Q-PAGE from cassette
Setting up gel/membrane sandwich for Western transfer
Recommendations/Tips for Gel Running
1. 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-PAGE
1. 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 Loading
1.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 is provided below. (Invitrogen® Mini Gel Tank 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 V | 180 V | 230 V*2 |
Running Time*1 | 60-75 mins | 35-50 mins | 25-40 mins |
Expected Current Initial (per gel) Final (per gel) |
70-80 mA 20-30 mA |
90-100 mA 35-45 mA |
130-140 mA 60-70 mA |
Expected temperature | 25-30°C | 25-35 °C | 35-45°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 supply.
Remove Q-PAGE™ Midi Gel from Cassette
Open 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 up notched plate and let gel stay on the front plate.
4.Use cassette opener to push through the slot in the cassette.
5.Carefully detach the gel from the bottom of gel
- Avoid diagonally peeling the gel from the corner.
- If necessary, cut well separators with gel remover
6.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.
*Activate PVDF membrane in methanol before soaking in transfer buffer.
**Prepare 6 filter papers for one gel/membrane sandwich.
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™ Midi Precast Gels to Invitrogen Mini Gel Tank Electrophoresis System
1. Place the Q-PAGE Midi Precast Gels with notched plate facing toward yourself. No extra adapter is needed.
2. Seat the gels on the bottom of Mini Gel Tank and close the cassette clamp.
3. Fill chambers with running buffer to the level of the fill line. Ensure gel wells are completely covered.
Adapting Q-PAGE™ Midi 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.
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Q-PAGE™ Precast Gel
Gel Type | Bis-Tris | TGN (Tris-Glycine-Novel) | ||||||
Buffer systems | MOPS and MES | Tris-Glycine (Laemmli) | ||||||
Features | Clear and sharp bands, high resolution | Quick running, clear bands | ||||||
Cassette size | Mini 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 system | Bio-Rad systems | Mini Gel Tank Xcell SureLock, Hoefer SE260 | Bio-Rad systems | Mini Gel Tank Xcell SureLock, Hoefer SE260 | ||||
Well format & Capacity | 12 wells, 25 μl/well | 15 wells, 22 μl/well | 12 wells, 40 μl/well | 15 wells, 28 μl/well | 12 wells, 25 μl/well | 15 wells, 22 μl/well | 12 wells, 40 μl/well | 15 wells, 28 μl/well |
Gel percentage/ Cat. No. | 8% | 8% | 8% | 8% | 10% | 10% | 10% | 10% |
QP2110 | QP2120 | QP3110 | QP3120 | QP4210 | QP4220 | QP5210 | QP5220 | |
12% | 12% | 12% | 12% | 4-15% | 4-15% | 4-15% | 4-15% | |
QP2310 | QP2320 | QP3310 | QP3320 | QP4510 | QP4520 | QP5510 | QP5520 | |
4-12% | 4-12% | 4-12% | 4-12% |
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QP2510 | QP2520 | QP3510 | QP3520 |
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ExcelBand™ Protein Markers
Ready-to-use— premixed with a loading buffer for direct loading, no need to boil
Broad range— 310 kDa to 5 kDa
Pre-stained bands — for monitoring protein separation during electrophoresis and Western blotting transferring efficiency on membrane
Enhanced bands— for quick reference
YesBlot™ Western Marker I
Ready-to-use — no need of mixing or heating before sample loading
Direct visualization — 10 IgG-binding proteins for direct visualization on Western blots
Pre-stained bands — 4 pre-stained proteins for monitoring protein separation during electrophoresis and Western blotting transferring efficiency on membrane
Wide range — 10 clear bands from 15 to 200 kDa for size estimation
Quick reference — two enhanced bands (30 and 80 kDa)
FluoroStain™ Protein Fluorescent Staining Dye
Compatible 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 staining
Substitution of the Coomassie Blue protein staining method
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求采纳为满意回答。
1、 缓冲溶液对反应物的测定没有干扰
2、缓冲组分的浓度为1:1
3、有足够的缓冲容量
4、缓冲溶液的PH应在所需范围内
5、组成缓冲溶液的弱碱PKB和弱酸PKA应接近或等于所需的POH值或PH值（PH+POH=14）
配制
只要知道缓冲对的PH值，和要配制的缓冲液的pH值（及要求的缓冲液总浓度），就能按公式计算[盐]和[酸]的量。这个算法涉及对数换算，较麻烦，前人为减少后人的计算麻烦，已为我们总结出pH值与缓冲液对离子用量的关系并列出了表格。只要我们知道要配制的缓冲液的pH，经查表便可计算出所用缓冲剂的比例和用量。例如配制500nmpH5.8浓度为0.1M磷酸缓冲液。
经查表知pH5.8浓度为0.2M Na2HPO48.0毫升，而0.2M Na2HPO492.0毫升。依此可推论出配制100ml0.1M的磷酸缓冲液需要0.1M Na2HPO48.0毫升，而0.1M Na2HPO4需要92.0毫升。
计算好后，按计算结果准确称好固态化学成分，放于烧杯中，加少量蒸馏水溶解，转移入50ml容量瓶，加蒸馏水至刻度，摇匀，就能得到所需的缓冲液。
各种缓冲溶液的配制，均按表格按比例混合，某些试剂，必须标定配成准确浓度才能进行，如醋酸、氢氧化钠等。另外，所有缓冲溶剂的配制计量都能从以上的算式准确获得。
在缓冲溶液中加入少量强酸或强碱，其溶液pH值变化不大，但若加入酸，碱的量多时，缓冲溶液就失去了它的缓冲作用。这说明它的缓冲能力是有一定限度的。
缓冲溶液的缓冲能力与组成缓冲溶液的组分浓度有关。0.1mol·L-1HAc和0.1mol· L-1NaAc组成的缓冲溶液，比0.01mol·L-1HAc和0.01mol·L-1NaAc的缓冲溶液缓冲能力大。关于这一点通过计算便可证实。但缓冲溶液组分的浓度不能太大，否则，不能忽视离子间的作用。
组成缓冲溶液的两组分的比值不为1∶1时，缓冲作用减小，缓冲能力降低，当c（盐）/c（酸）为1∶1时△pH最小，缓冲能力大。不论对于酸或碱都有较大的缓冲作用。缓冲溶液的pH值可用下式计算：
此时缓冲能力大。缓冲组分的比值离1∶1愈远，缓冲能力愈小，甚至不能起缓冲作用。对于任何缓冲体系，存在有效缓冲范围，这个范围大致在pKaφ（或pKbφ）两侧各一个pH单位之内。
弱酸及其盐（弱酸及其共轭碱）体系pH=pKaφ±1
弱碱及其盐（弱碱及其共轭酸）体系pOH=pKbφ±1
例如HAc的pKaφ为4.76，所以用HAc和NaAc适宜于配制pH为3.76～5.76的缓冲溶液，在这个范围内有较大的缓冲作用。配制pH=4.76的缓冲溶液时缓冲能力最大，此时（c（HAc）/c（NaAc）=1。
制备
为了配制一定pH的缓冲溶液，首先选定一个弱酸，它的pKaφ尽可能接近所需配制的缓冲溶液的pH值，然后计算酸与碱的浓度比，根据此浓度比便可配制所需缓冲溶液。
以上主要以弱酸及其盐组成的缓冲溶液为例说明它的作用原理、pH计算和配制方法。对于弱碱及其盐组成的缓冲溶液可采用相同的方法。
缓冲溶液在物质分离和成分分析等方面应用广泛，如鉴定Mg2+ 离子时，可用下面的反应：
白色磷酸铵镁沉淀溶于酸，故反应需在碱性溶液中进行，但碱性太强，可能生成白色Mg（OH）2沉淀，所以反应的pH值需控制在一定范围内，因此利用NH3·H2O和NH4Cl组成的缓冲溶液，保持溶液的pH值条件下，进行上述反应。
常用缓冲液配制
枸橼酸-磷酸氢二钠
甲液:取枸橼酸21g或无水枸橼酸19.2g,加水使溶解成1000ml，置冰箱内保存。
乙液：取磷酸氢二钠71.63g，加水使溶解成1000ml。
取上述甲液61.45ml与乙液38.55ml，混合，摇匀，即得。
氨－氯化铵缓冲液
取氯化铵1.07g，加水使溶解成100ml， 再加稀氨溶液(1→30)调节pH值至8.0，即得。
氨－氯化铵缓冲液
取氯化铵5.4g，加水20ml溶解后，加浓氨溶液35ml，再加水稀释至100ml，即得。
醋酸－醋酸钠缓冲液
取无水醋酸钠20g，加水300ml溶解后,加溴酚蓝指示液1ml及冰醋酸60～80ml，至溶液从蓝色转变为纯绿色，再加水稀释至1000ml，即得。
醋酸－醋酸钠缓冲液
取醋酸钠18g,加冰醋酸9.8ml,再加水稀释至1000ml，即得。
醋酸－醋酸钠缓冲液
取醋酸钠54.6g，加1mol/L醋酸溶液20ml溶解后，加水稀释至500ml，即得。
醋酸－醋酸铵缓冲液
取醋酸铵7.7g，加水50ml溶解后，加冰醋酸6ml与适量的水使成100ml，即得。
醋酸－醋酸铵缓冲液
取醋酸铵77g，加水约200ml使溶解，加冰醋酸57ml,再加水至1000ml，即得。
醋酸－醋酸铵缓冲液
取醋酸铵100g，加水300ml使溶解，加冰醋酸7ml,摇匀，即得。
磷酸盐缓冲液
取0.2mol/L磷酸二氢钾溶液250ml,加0.2mol/L氢氧化钠溶液118ml，用水稀释至1000ml，即得。
当往某些溶液中加入一定量的酸和碱时，有阻碍溶液pH变化的作用，称为缓冲作用，这样的溶液叫做缓冲溶液。弱酸及其盐的混合溶液（如HOAc与NaOAc），弱碱及其盐的混合溶液（如NH3·H2O与NH4Cl）等都是缓冲溶液。
参考资料：http://baike.baidu.com/view/901429.htm
