本产品适用于Tris-甘氨酸电泳体系，采用上层胶和下层胶的预混配方，只需加入改良型促凝剂即可凝胶，灌入下层胶后，无需等待凝胶，直接灌入上层胶即可，简便快捷。所配的上层胶带有颜色(红色、蓝色或绿色)，点样孔清晰易辨，方便点样。三种颜色设计，可用于区分含不同样品的凝胶。本试剂盒灌制的凝胶也可用于非变性PAGE凝胶电泳。
本产品配套提供改良型促凝剂 ，其具有更好的稳定性和催化效能，配胶过程中无需额外添加TEMED。为方便操作，已开盖的改良型促凝剂可置于4℃保存至少三个月。
本产品可制胶数量：125块（0.75mm胶）或＞90块（1.00mm胶）或＞60块（1.50mm胶）。
| 货号 | 上层胶浓度
| 下层胶浓度
|
T1491535-01
| 4.5%
| 6%
|
T1491535-02
| 4.5%
| 7.5%
|
T1491535-03
| 4.5%
| 10%
|
T1491535-04
| 4.5%
| 12.5%
|
T1491535-05
| 4.5%
| 15%
|
T1491535
| Component
| 125gels (0.75mm胶)
| Storage
|
T1491535A
| 上层胶溶液 (2×)
| 80 mL | 2-8℃ |
T1491535B
| 彩色上层胶缓冲液 (2×)
| 80 mL
| 2-8℃
|
T1491535C
| 下层胶溶液 (2×)
| 250 mL
| 2-8℃
|
T1491535D
| 下层胶缓冲液 (2×)
| 250 mL
| 2-8℃
|
T1491535E
| 改良型促凝剂
| 8 mL
| -20℃ |
产品特点
1、一步法灌胶——灌制下层胶后直接注入上层胶，无需液封；
2、操作简单快速——无需计算所需溶液量，无需稀释；
3、彩色上层胶——可制备红蓝绿三种颜色的上层胶，为点样和区分不同凝胶提供便利；
4、避免异味——无需使用TEMED，避免恶臭气味；
5、条带清晰——尤其小分子蛋白质条带比在传统凝胶中更清晰。
制胶流程(以一块0.75/1.0/1.5mm的mini胶为例)
1、取等体积下层胶溶液和下层胶缓冲液，各2.0/2.7/4.0mL，混匀；
2、取等体积上层胶溶液和彩色上层胶缓冲液，各0.5/0.75/1.0mL，混匀；
注意：由于染料的特殊理化性质，使用前请摇匀。
3、向步骤1的混合溶液中加入40/60/80μL的改良型促凝剂，轻轻混匀，将混匀后的溶液注入制胶玻璃板中，使液面和短玻璃板上沿之间的距离比梳齿长0.5cm即可；
注意：
① 此溶液为过量，请勿全部注入；
② 加入改良型促凝剂后，需轻柔混匀，防止过多氧气混入胶溶液，抑制凝胶聚合。
4、向步骤2的混合溶液中加入10/15/20μL的改良型促凝剂，轻轻混匀，无需等待下层胶凝固，即可将混匀后的溶液轻缓注入制胶玻璃板中，插入梳齿；
注意：
① 灌注上层胶溶液一定要轻缓，避免将上层胶溶液冲入下层胶，上层胶溶液注入后，轻轻振动制胶架，即可使上下层胶分界线平齐；
② 加入改良型促凝剂后，需轻柔混匀，防止过多氧气混入胶溶液，抑制凝胶聚合。
5、待胶凝固后（约15min），拔去梳齿即可用于电泳。
注意：
① 请尽量使用新鲜配制的电泳缓冲液；
② 胶凝固后上下层胶分界线平整度略弱于传统方法配的胶，但对后续电泳没有影响。
下层胶配方：
| 凝胶厚度 | 下层胶溶液 | 下层胶缓冲液
| 改良型促凝剂 |
| 0.75mm | 2.0 mL | 2.0 mL
| 40 μL
|
| 1.00mm | 2.7 mL
| 2.7 mL
| 60 μL
|
| 1.50mm | 4.0 mL
| 4.0 mL
| 80 μL |
上层胶配方：
| 凝胶厚度 | 上层胶溶液 | 上层胶缓冲液
| 改良型促凝剂 |
| 0.75mm | 0.5 mL | 0.5 mL
| 10 μL
|
| 1.00mm | 0.75 mL
| 0.75 mL
| 15 μL
|
| 1.50mm | 1.0 mL
| 1.0 mL
| 20 μL |
注意事项
1、本产品制备出的凝胶其上层胶对样品没有浓缩效应，与预制胶类似，但与传统PAGE胶相比，对蛋白条带分离效果更好， 小分子量蛋白(比如10kDa)也可以清晰地分离开，且蛋白条带更窄更锐利；
2、不同浓度试剂盒各组分请勿混用，否则会影响制胶及电泳效果；
3、改良型促凝剂的使用量仅作参考，实际用量可根据个人实验习惯和经验调整。加入较多量的促凝剂可加速凝胶，反之亦然；
4、凝胶速度与温度有显著的正相关性。同等条件下，温度越高，凝胶速度越快，室温过高时建议适当减小改良型促凝剂的用量；相反，如果室温较低，可适当延长凝胶时间；
5、本产品已加入适量TEMED的替代品，如需进一步加速凝胶，临配胶前可按需补充适量TEMED；
6、在配胶之前，使胶溶液及缓冲液平衡到室温(如室温放置几分钟)，可有效避免凝胶中气泡的形成；
7、推荐电泳条件为：150V，约60min（或200V，约45min）；
8、为了您的安全和健康，请穿实验服并戴一次性手套操作；
9、本产品仅限科研使用。
凝胶浓度选择参考
下图为Tris-Gly缓冲系统中，蛋白分子量标准（10-250kDa），含有11条蛋白条带在不同浓度的SDS-PAGE凝胶中的分离示意图。因温度、pH值等因素不同，实际分离情况会略有出入，本图仅供参考。
This product is designed for the Tris-glycine electrophoresis system. It features pre-mixed formulations for both stacking (upper) and separating (lower) gels. Polymerization is achieved simply by adding the Modified Polymerizer—no additional TEMED is required. After pouring the lower gel, the stacking gel can be added immediately without waiting for polymerization. The prepared stacking gel is colored (red, blue, or green) for clear well identification and easy sample loading. The three color options allow easy distinction between gels loaded with different samples. Gels prepared with this kit are also suitable for native PAGE.
The included Modified Polymerizer offers improved stability and catalytic efficiency. Once opened, it can be stored at 4°C for at least three months.
Gel Yield: 125 gels (0.75 mm) or 90 gels (1.00 mm) or 60 gels (1.50 mm)
Product Code
| Stacking Gel
| Separating Gel
|
T1491535-01
| 4.5%
| 6%
|
T1491535-02
| 4.5%
| 7.5%
|
T1491535-03
| 4.5%
| 10%
|
T1491535-04
| 4.5%
| 12.5%
|
T1491535-05
| 4.5%
| 15%
|
T1491535
| Component
| 125gels (0.75mm)
| Storage
|
T1491535A
| Stacking Gel Solution (2×)
| 80 mL | 2-8℃ |
T1491535B
| Colored Stacking Buffer (2×)
| 80 mL
| 2-8℃
|
T1491535C
| Separating Gel Solution (2×)
| 250 mL
| 2-8℃
|
T1491535D
| Separating Gel Buffer (2×)
| 250 mL
| 2-8℃
|
T1491535E
| Modified Polymerizer
| 8 mL
| -20℃ |
Features
1.One-Step Casting: The stacking gel can be poured directly after the separating gel without isopropanol or water sealing.
2.Easy & Fast Operation: No need to calculate volumes or perform dilutions.
3.Colored Stacking Gel: Available in red, blue, or green for easy well identification and sample distinction.
4.Odor-Free: TEMED is not required, eliminating unpleasant odors.
5.Sharp Bands: Especially improves resolution for low-molecular-weight proteins compared to traditional gels.
Gel Preparation Protocol (for one 0.75/1.0/1.5 mm mini gel)
1.Mix equal volumes of Separating Gel Solution and Separating Gel Buffer (2.0/2.7/4.0 mL each).
2.Mix equal volumes of Stacking Gel Solution and Colored Stacking Buffer (0.5/0.75/1.0 mL each).
Note: Due to the properties of the dye, mix well before use.
3.Add 40/60/80 µL of Modified Polymerizer to the separating gel mixture. Mix gently and pour into the gel cassette until the level is 0.5 cm above the top of the short plate.
Note:
· This volume is excessive; do not use all.
· Avoid vigorous mixing to prevent oxygen incorporation that may inhibit polymerization.
4.Add 10/15/20 µL of Modified Polymerizer to the stacking gel mixture. Mix gently. Without waiting for the lower gel to set, slowly pipette the stacking gel mixture onto the polymerizing separating gel. Insert the comb.
Note:
· Pour slowly to avoid disturbing the lower gel. Gently tap the cassette to level the interface.
· Mix gently to avoid introducing oxygen.
5.After polymerization (~15 minutes), remove the comb. The gel is ready for electrophoresis.
Note:
·Fresh electrophoresis buffer is recommended.
· The interface may appear less smooth than in traditionally cast gels but does not affect electrophoresis.
Separating Gel
Gel Thickness
| Separating Gel Solution
| Separating Gel Buffer
| Modified Polymerizer
|
| 0.75mm | 2.0 mL | 2.0 mL
| 40 μL
|
| 1.00mm | 2.7 mL
| 2.7 mL
| 60 μL
|
| 1.50mm | 4.0 mL
| 4.0 mL
| 80 μL |
Stacking Gel
Gel Thickness
| Stacking Gel Solution | Colored Stacking Buffer
| Modified Polymerizer
|
| 0.75mm | 0.5 mL | 0.5 mL
| 10 μL
|
| 1.00mm | 0.75 mL
| 0.75 mL
| 15 μL
|
| 1.50mm | 1.0 mL
| 1.0 mL
| 20 μL |
Precautions
1.Gels made with this kit do not provide sample stacking like traditional Laemmli gels but offer improved protein separation—especially for low-MW proteins (e.g., 10 kDa)—with sharper and narrower bands.
2.Do not mix components from kits of different concentrations.
3.The amount of Modified Polymerizer is a guideline and can be adjusted based on experience. More accelerator speeds up polymerization; less slows it down.
4.Polymerization rate is temperature-dependent. Reduce accelerator amount at high temperatures; extend polymerization time at low temperatures.
5.If faster polymerization is needed, additional TEMED may be added right before use.
6.Bring all solutions to room temperature before use to minimize bubble formation.
7.Recommended electrophoresis conditions: 150 V for ~60 min or 200 V for ~45 min.
8.Wear a lab coat and disposable gloves during operation.
9.For research use only.
J2609474
危险品化学品经营许可证（带存储）