![SMOBIO/[QP4220] Q-PAGE™ TGN Precast Gel (Mini, 15 wells, 10%), 10 gels/Mini, 15 wells, 10%), 10 gels</span>
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Description
Q-PAGE™ TGN (Tris-Glycine Novel) Precast Gels are ready-to-use acrylamide gels for SDS-PAGE running in Tris-Glycine buffer system. With unique formula, Q-PAGE™ TGN Precast Gels perform enhanced speed, better separation, and longer shelf life as compared with conventional Laemmli Tris-HCl gels. The protein migration patterns in Q-PAGE™ TGN series, however, are similar with typical Laemmli Tris-HCl gels, and thus Q-PAGE™ TGN Precast Gels are compatible to traditional SDS-PAGE and subsequent analyses.
Q-PAGE™ TGN Precast Gels are available in gradient (4 to 15%) and fixed (10%) 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 (QP4XXX) Gels are suitable for Bio-Rad® and other systems. Q-PAGE™ Midi (QP5XXX) 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
Labeled warning sign and green tape as reminder
Enhanced gel performance:
Enhanced gel electrophoresis speed
Better band separation
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.
Technical
Quick running, clear bands
Q-PAGE™ TGN Precast Gel can separate protein in 19 minutes using 300 V.
QP4220 Specifications
Gel | TGN (Tris-Glycine-Novel) | |
Buffer systems | Tris-Glycine (Laemmli) | |
Features | Quick running, clear bands | |
Cassette size | Mini Gel (10 X 8.3 cm) | |
Gel dimensions
| 8.1 x 7.4 x 0.1 cm (W x L x thickness) cm | |
Electrophoresis system | Bio-Rad systems | |
Well format & Capacity | 15 wells, 22 μl/well | |
Gel percentage | 10 % | |
Accessory tray | Production description Tip card Gel remover Cassette opener | |
Manual
Manual_Q-PAGE™ TGN Precast Gel, Mini
SDS
SDS_Q-PAGE™ Precast Gel
Migration pattern
Setting Up and Running Q-PAGE™ Mini Precast Gel
Tips for removing Q-PAGE Mini Gel 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. Rinse the wells before sample loading.
4. Try 200 V first, and optimize the voltage and running time if needed. Do not set voltage lower than 100 V.
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 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.
| 150 V | 200 V*2 | 250 V*3 | 300 V*3 |
Running Time*1 | 40-60 mins | 30-40 mins | 25-35 mins | 15-25 mins |
Expected Current Initial (per gel) Final (per gel) |
40-50 mA 10-20 mA |
50-60 mA 25-30 mA |
80-90 mA 35-40 mA |
90-100 mA 40-50 mA |
Expected temperature | 25-30°C | 25-40 °C | 25-40°C | 25-40°C |
*1 Set voltage higher than 100 V is recommended
*2 Try 200 V first, and optimize the voltage and running time if needed.
*3For higher voltage conditions, please use fresh running buffer for inner and outer chambers
*4 Running time varies depending on gel percentage, running buffer, temperature, and power supply.
Remove Q-PAGE™ 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 two plates apart from the top of cassette.
4.Carefully detach the gel either from the bottom or the top side of the cassette.
-Avoid diagonally peeling the gel from the corner.
-Use water to help gel detachment if needed.
5.Gently remove the gel for further staining or Western blotting.
Troubleshooting Guidelines | ||
Problem | Possible Cause | Suggested Solution |
Well deformation | Pull one side of comb out of cassette. | Smoothly pull the comb straight out of the cassette. |
Bubbles between gel and cassette | Gel has been frozen or stored at wrong temperature. | Store Q-PAGE Precast Gels at 4°C. |
Buffer leaking from the inner chamber | Untight assembly of gels to the electrode modules | Reassemble 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 wells | Rinse the gel wells with ddH2O or 1X running bufferbefore loading. |
Insufficient sample buffer | Use more sample buffer to prepare samples. | |
Current is zero and sample do not migrate into gel | Tape at bottom of gel not removed | Remove tape |
Gels run faster or more slowly than expected. | Incorrect running buffer | Check buffer composition. Use fresh 1X running buffer for inner chamber. |
Crooked bands at middle or bottom of gel | Gel has been frozen or stored at wrong temperature. | Store Q-PAGE Precast Gels at 4°C. |
Incorrect running buffer | Check 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 chamber | Check assembly of gels into the electrode modules. |
Excessive heating of gel | Check buffer composition. Or dilute running bufferto 0.5-0.75X. Do not exceed recommended running conditions. | |
Insufficient buffer in inner or outer buffer chamber | Fill inner and outer chambers to completely covergel wells. | |
Poor resolution or fuzzy bands | Excessive heating of gel | Check buffer composition. Do not exceed recommended running conditions. |
Incorrect running buffer | Check buffer composition. | |
Bands are missing on the membrane after Westerntransferring. | Proteins move in the wrong direction | Check 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/membranesandwich Remove 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 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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HT培养基添加剂(HT Supplement)为粉剂，经照射灭菌。每个包装可配制成10ml浓缩液（50X），用于杂交瘤细胞的筛选。
1、常用玻璃仪器的准备制备培养基所用的吸管、锥形瓶、毛细吸管、平皿等玻璃仪器用前要用肥皂水洗刷后用清水冲洗干净，晾干后备用。
（1）平皿用纸或布包好，或装在金属盒内，于121℃高压蒸汽菌3min后烘干，备用。
（2）试管管口用棉花塞或硅氟塑料试管塞塞好，再用布或报纸包扎好，121℃高压蒸汽灭菌20℃后烘干，备用。
（3）吸管及滴管先用少许棉花塞于吸口端（防止污染物吸出，或橡皮帽内的气体污染），然后用纸包后或装入金属吸管筒内，于121℃高压蒸汽灭菌20min后烘干，备用。其他玻璃器皿均应按上法处理，也应装金属筒内160℃干热灭菌2h后，备用。
2、培养基的制备及储存
（1）溶化：一般应放在玻璃器皿或搪瓷齐内。加入蒸馏水，隔水加热以促其溶解，加热时应经常搅拌，防止焦结，待其溶解后补足水分。
在使用干燥培养基时，先将蒸馏水按定量加入容器中，然后称取一定量培养基干粉放入水中，静置10—15min，搅拌，振荡，或延长时间以促进溶解，一般不要热，必要时加热，但时间不宜过长，温度不宜过高，避免某些营养成分被破坏。
（2）校正酸碱度（调节pH）：培养基必须有适当的pH。因此测定pH是培养基配制过程中的重要步骤之一，测定pH的标准温度为25士2℃。调节pH可用无菌的1mol/l氢氧化钠溶液或10%碳酸钠溶液、1mol/l盐酸溶液。当调节缓冲液的pH时，宜用6mol/l磷酸或10mol/l氢氧化钾溶液。灭菌后的pH会比灭菌前的pH升高或降低，一般高压灭菌前培养基的pH会比最终pH调高0.2左右，灭菌后基本合适。因此对培养基、缓冲液、试剂在灭菌前后的pH均进行测定，并记下每次pH的测定结果，有助于积累数据考察每种培养基、缓冲液、试剂对灭菌程序的耐受性，从而指导灭菌前pH的调节。干燥培养基一般已校正过pH，用时也必须再验证。测定时，一般用指示剂滴入培养基观察其颜色的变化，或用pH计校正，如与所需pH不符，可用以上的酸或碱液加以校正。调整pH后要加热过滤，使培养基澄清。( 更多质量检测、分析测试、化学计量、标准物质相关技术资料请参考中国标准物质 www.rmhot.com)
（3）培养基的分装：大批量配制时使用的自动分配器须经校准和确认。每次使用前后，均要对分配器的管道系统进行冲洗，在分装无菌培养基前，则要采用无菌硅胶管。
固体培养基一般分装在250ml、500ml锥形瓶中，高压灭菌后根据需要分装平皿或试管。
平皿：倾注平皿，应在无菌室中放置3—4h，如用塑料平皿须在35℃培养箱 中倒置30—60min。让水蒸汽自然蒸发。
斜面：制备低层斜面分装于试管，约占容积1/5，灭菌后趁热斜放在细玻璃棒和木杆上，使成斜度，分装使注意管口和瓶塞上勿沾有培养基，如沾有培养基应用布抹去，以避免污染。
高层斜面：制备高层斜面分装于试管，约占试管容积的1/3，灭菌后趁热放置成高层短斜面，待其凝固后应用。
分装好的培养基或缓冲液等及时密封后必须在配制当天（2h内最佳）进行灭菌处理。液体、半固体培养基一般在高压灭前分装，约装试管容积的1/3，在灭菌后还要加其他成分的液体、半固体培养基，灭菌后再分装于灭菌试管或锥形瓶中。
培养基的灭菌：培养基的灭菌多采用高压蒸汽灭菌，各种培养基的灭菌时间和压力，按其成分不同而定。普通培养基采用121℃、103.42kPa灭菌15min，但容器和装量较大时，应延长至20min。含糖培养基115℃、68.85kPa灭菌30min。含糖、血清、 鸡蛋等培养基可用流动蒸汽及血清凝固器80—100℃、30min，连续3d，间歇灭菌。血清或组织液，采用低热56—58水浴1h，连续5—6d灭菌，一些遇高温即被破坏的物质如尿素、腹水等，可用细菌过滤器，过滤除菌。高压灭菌应严格按照操作规程执行，必须逐渐加热，彻底排除灭菌器内的空气，再逐渐升压保持预定的压力和时间，达到全部杀灭微生物。以上的灭菌时间和温度要经过验证确认，如不同类型培养基混合一起灭菌时宜采用115℃、68.85kPa灭菌30min为好。
通过无菌技术为测试制备的每一批培养基其PH在放冷至室温（25°）后，都应测定。一个扁平的pH计用于培养基表面pH值的测定，浸入式的pH计用于液体的测定。各供应商提供的培养基的pH应该在规定的±0.2的范围内，除非通过验证得到更宽的范围
注意：按配方计算培养基中各种成分的用量→称量,（一般动作要迅速一些,因为其中可能有些成分容易吸潮）→溶化（将称好的各种成分溶解在水中,如果是固体培养基,需要使用凝固剂,如琼脂等,熔化时,注意搅拌,防止糊底）→调pH→灭菌（高压蒸汽灭菌）→倒平板
1）确认培养基的成分,并精确称量；
2）加入各个成分的顺序；
3）准确调pH；
4) 适当的灭菌方法；
5）无菌条件下分装.
各位大神你们好：
我们是医疗器械工厂，只需配制环境监测和水检测的培养基，但是根据2015版药典规定，1105，141页有一句
而在9203，289页
“商品化的成品培养基除了应附有处方和使用说明外，还应注明有效期、贮藏条件、适用性检查试验的质控菌和用途”。
是否可以理解为只要供应商提供以上信息，使用厂家就可以不用做培养基适用性试验了？
如果购买的商品化的成品培养基不做培养基适用性试验，那么医疗器械的微生物检验将大大减少工作量了。
