iBlot 7-分钟干式蛋白转印系统

tel: 400-6699-117 转 9566

赛默飞蛋白印迹仪/转印仪, 全新的iBlot® 7-分钟转印系统可在7分钟内对聚丙烯酰胺凝胶中的蛋白质进行转印。 iBlot®蛋白质印迹系统可高效转......

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一般可在7分钟或更短的时间内实现蛋白的完全转移

Comparison of the three types of electrotransfer apparatus

We have compared the iBlot Dry Blotting System to the conventional semi-dry transfer and wet western transfer systems. Our results demonstrate that the iBlot dry blotting method provides superior immunodetection sensitivity—that is, transfer quality—compared to either semi-dry or wet transfer methods (Figures 1 and 2). In addition, the iBlot Dry Blotting System is faster and more efficient than other blotting methods (Table 1).

Figure 1. High transfer efficiencies achieved using the iBlot Dry Blotting System. (A) iBlot dry transfer to nitrocellulose and (B) semi-dry transfer to nitrocellulose of NuPAGE Novex 12% Bis-Tris mini gels. Lanes 1–6: 0.0625 µg, 0.125 µg, 0.25 µg, 0.5 µg, 1.0 µg, and 2.0 µg SW480 human colon cancer cell lysate; lanes 7 and 12: 5 µL SeeBlue Plus2 Pre-Stained Protein Standard; lanes 8–11: 0.5 µL, 1.0 µL, 2.0 µL, 4.0 µL MagicMark XP Western Protein Standard.

Figure 2. High transfer quality achieved using the iBlot® Dry Blotting System. (A) iBlot dry transfer to nitrocellulose and (B) wet (tank) transfer to nitrocellulose of NuPAGE Novex 12% Bis-Tris mini gels. Lanes 2–7: 0.0625 µg, 0.125 µg, 0.25 µg, 0.5 µg, 1.0 µg, and 2.0 µg SW480 human colon cancer cell lysate; lanes 1 and 8: 5 µL SeeBlue Plus2 Pre-stained protein standards; lanes 9–12; 0.5 µL, 1.0 µL, 2.0 µL, 4.0 µL MagicMark XP Western Protein Standard.

Table 1. Comparison of elapsed time for protein transfer with the iBlot Dry Blotting System to other blotting methods.

	iBlot Dry Blotting System	Semi-dry transfer	Wet or semi-wet transfer
Buffer preparation	0 minutes	30 minutes	30 minutes
Soaking gel in transfer buffer	0 minutes	20 minutes	0 minutes
Assembling layers	2 minutes	10 minutes	10 minutes
Transfer	7 minutes	45–90 minutes	1–3 hr
Cleanup	0 minutes	10 minutes	10 minutes
Total elapsed time	9 minutes	1 hr, 55 min– 2 hr, 40 min	1 hr, 50 min– 3 hr, 50 min
Time saved with the iBlot Dry Blotting System	—	1 hr, 45 min– 2 hr, 30 min	1 hr, 40 min– 3 hr, 40 min

Method overview of the three approaches

iBlot Dry Blotting System	Semi-dry transfer	Wet or semi-wet transfer
(Enlarge)	(Enlarge)	(Enlarge)
Preassembled stacks ready for protein transfer containing electrodes, buffer matrices, and PVDF or NC membrane	Transfer stack (both top and bottom) composed of sponge and filter paper, soaked in buffer	Transfer stack composed of sponge and filter paper, soaked in a tank filled with buffer

Comparison of properties

	iBlot Dry Blotting System	Semi-dry transfer	Wet or semi-wet transfer
Transfer buffer required?	No	100–250 mL, or just enough to construct a bubble-free sandwich	1–1.5 L, or enough to fill the transfer tank
Transfer time	7 min, plus 2 min transfer preparation	45–90 min, plus 70 min preparation and assembly	1 hr–overnight, plus 50 min preparation and assembly
Transfer quality	Reproducible and good transfer quality for proteins between 11 and 220 kDa:* Transfer protocols optimized for immunodetection, e.g., iBlot® system–transferred membranes exhibit higher immunodetection sensitivity, using chromogenic and chemiluminescent procedures	Variable and inefficient transfer quality: Reduced buffer capacity limits transfer time, especially for mid- to large molecular weight proteins Small molecular weight proteins may be transferred through membrane and onto filter paper below Membrane and filter paper MUST be cut to exact gel size, otherwise current will short-circuit around the edge of the gel	Reliable and good transfer quality: Increase temperature during blotting, unless buffer is mixed and cooled during blotting keeps current relatively constant High-current power source is typically required for 1–2 hour transfer
Supplemental equipment required	None	External power supply	External power supply
Post-transfer requirement	None	Wet-soaking filter paper for clean-up Salt deposits on electrodes require regular maintenance	Large amount of hazardous buffer to discard Wet-soaking sponges for clean-up Salt deposits on electrodes require regular maintenance