iBind™ Solution Kit, 4 kits - FAQs

除了iBind Flex卡片的宽度是iBind卡片的1.7倍之外，iBind Flex卡片背面还印有对齐线，它们会在加入溶液时出现，有助于正确水平摆放多张小型或垂直剪切膜条。

两种标准和荧光检测溶液的配方是相同的。唯一的不同点是产品供应量不同，iBind Flex溶液试剂盒足够供10块中型、20块小型或60块垂直剪切膜条使用。

当使用iBind蛋白质免疫印迹系统和LI-COR Odyssey成像系统时，请采用使用手册（https://assets.thermofisher.com/TFS-Assets/LSG/manuals/ibind_man.pdf）第13页的实验方案。我们不建议使用LI-COR溶液。

如果PVDF膜是干燥的，则在浸泡于iBind溶液/ iBind荧光检测（FD）溶液之前，需要使用100%甲醇预活化，并用蒸馏水润洗。刚刚转印完毕并转移至在iBind溶液/ iBind荧光检测（FD）溶液中的PVDF膜无需活化。

每次使用后，用流水冲洗iBind蛋白质印迹设备，在下一次使用前将设备晾干。不建议使用去污剂，因为残留的去污剂可能会干扰结果。

为延长iBind蛋白质印迹设备弹簧的使用寿命，保存设备时应将弹簧锁解锁，并打开盖子，如使用手册(https://tools.thermofisher.com/content/sfs/manuals/ibind_man.pdf)第23页所示。

将膜置于iBind溶液/ iBind荧光检测（FD）溶液中浸泡至制备抗体稀释液之前，足以完成对膜的封闭。不需要将膜在封闭液中封闭过夜。加入一抗之前，膜会在iBind溶液/ iBind荧光检测（FD）溶液中额外受到封闭。

可以，小于9 cm x 9 cm的膜可以用于iBind蛋白质免疫印迹系统。建议将膜条或更小的印迹膜放在设备中心。使用尺寸较大的膜时，必须确保膜的任何部分都没有与iBind卡片上的膜组接触。

我们不建议使用您自己的溶液。iBind溶液和iBind荧光检测（FD）溶液具有特殊的粘稠度，并且针对iBind蛋白质免疫印迹系统的顺序侧流原理进行了优化。我们不能对任何其他溶液的性能进行保证。

iBind蛋白质免疫印迹系统（货号SLF1000）由以下组件构成：

•iBind蛋白质印迹设备
•iBind印迹滚筒
•iBind观察窗盖

下列物品需单独购买：
•iBind卡片（货号SLF1010）
•iBind溶液试剂盒（货号SLF1020），制备用于化学发光或发光检测的封闭、稀释和洗涤缓冲液
•iBind荧光检测溶液试剂盒（货号SLF1019），制备用于荧光检测的封闭、稀释和洗涤缓冲液
•Invitrogen AP小鼠化学发光检测试剂盒（货号SLF1021）或Invitrogen AP兔化学发光检测试剂盒（货号SLF1022）

iBind蛋白质免疫印迹系统是一台自动化免疫检测设备。该设备无需外接电源，利用机械压力使免疫检测试剂发生顺序侧流（SLF），从而自动完成免疫印迹检测中所有的封闭、抗体孵育和洗涤步骤。

有两种iBind蛋白质免疫印迹系统可供选择：包括1）传统iBind蛋白质印迹设备，可容纳1块小型印迹膜，以及2）iBind Flex蛋白质印迹设备，可同时容纳1或2块小型印迹膜、1块中型印迹膜或最多6块垂直剪切膜条（用于不同抗体）。

To strip and reprobe your blot processed using the iBind Western Device or iBind Flex Western Device, first strip the blot outside the system using your preferred protocol. After the stripping protocol, wash the blot in distilled water for 5 min. Reblocking is unnecessary as the iBind Solution will perform this step during reprobing. Finally, reprobe with new antibodies using the iBind Western Device or iBind Flex Western Device, starting with the membrane preparation step.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Yes, for each midi blot, mini blot, or vertically cut strip, prepare appropriate primary antibodies together in a single tube of iBind FD/ iBind Flex FD Solution, Prepare appropriate secondary antibodies together in a different tube of iBind FD/ iBind Flex FD Solution. Do not mix primary and secondary antibodies prior to loading in the device. All primary and secondary antibodies should be diluted according to the recommendations outlined in the user manual or quick reference card. When detecting several targets of the same species, the same concentration of secondary antibody can be used as when detecting one target. There is no need to increase the concentration for each additional target. Load multiplexed antibodies into the device as normal.
Caution: Consider cross-reactivity of secondary antibodies when multiplexing (e.g., rabbit anti-goat IgG and goat anti-mouse IgG are likely to cross-react).

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Yes, both the iBind Western Device and iBind Flex Western Device work well with Alexa Fluor Plus conjugated secondary antibodies.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

When firmly rolling, the iBind/iBind Flex Card will slightly dip in the membrane area, causing the top and bottom of the card to lift. See the iBind/iBind Flex user manual for additional details.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Besides being ~1.7 times as wide, the iBind Flex Card also includes alignment lines printed on the back that appear when solution is added to the card to help in proper horizontal placement of multiple mini or vertically cut strip blots.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

The formulations are identical for both the standard and Fluorescent Detection (FD) solutions. The only difference is in the amount supplied, so the iBind Flex Solution Kits are sufficient for 10 midi, 20 mini, or 60 vertically cut strip blots.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Please use the protocol mentioned on Page 13 of the manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/ibind_man.pdf) when using the iBind Western System in conjunction with the LI-COR Odyssey Imaging System. We do not recommend using the LI-COR solution.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

If the PVDF membrane has dried up, it is important to pre-activate it with 100% methanol and then rinse in distilled water before immersing it in iBind Solution/iBind Fluorescent Detection (FD) Solution. It is not necessary to activate a PVDF membrane that has just come out of the transfer and moved into iBind Solution/iBind Fluorescent Detection (FD) Solution.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Rinse the iBind Western Device under running water after each use and allow the device to dry before additional usage. No use of detergent is recommended as any residual detergent may interfere with results. To maximize the life of the springs in the iBind Western Device, store the device with latch unlocked, and the lid open as shown on Page 23 of the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_man.pdf).

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

It is sufficient to block the membrane in iBind Solution/ iBind Fluorescent Detection (FD) Solution for the time prior to making antibody dilutions. It is not necessary to block the membrane overnight in the blocking solution. The membrane will be subjected to additional blocking in the iBind Solution/ iBind Fluorescent Detection (FD) Solution before primary antibody reaches the membrane.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

Yes, membranes that are smaller than 9 cm x 9 cm can be used with the iBind Western device. It is recommended to place the strips or smaller blots centrally. When using larger membranes, care must be taken to ensure that no part of the membrane is in contact with the stack on the iBind card.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

We do not recommend using your own solutions. The iBind Solution and iBind Fluorescent Detection (FD) Solution have specific viscosity and are optimized for the sequential lateral flow that is the principle of the iBind Western device. We cannot guarantee the performance with any other solutions.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

The original iBind Western System (Cat. No. SLF1000) consists of:
- iBind Western Device
- iBind Blotting Roller
- iBind Window Cover

You would need to purchase the following items separately:
- iBind Cards (Cat. No. SLF1010)
- iBind Solution Kit (Cat. No. SLF1020) for preparing blocking, dilution, and washing buffers for chemiluminescent or chromogenic detection
- iBind Fluorescent Detection Solution Kit (Cat. No. SLF1019) for preparing blocking, dilution, and washing buffers for fluorescent detection
- Invitrogen AP Mouse Chemiluminescent Detection Kit (Cat. No. SLF1021) or Invitrogen AP Rabbit Chemiluminescent Detection Kit (Cat. No. SLF1022)

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

iBind Western Systems are benchtop devices that automate immunodetection steps. These devices use no external power source, and rely on mechanical pressure to generate sequential lateral flow (SLF) of immunodetection reagents to perform hands-free blocking, antibody binding, and washes for western detection workflows.

Two iBind Western Systems are available:
i) containing the original iBind Western Device, which accommodates one mini blot at a time
ii) containing the iBind Flex Western Device, which accommodates one or two mini blots, one midi blot, or up to six vertically cut membrane strips (for separate antibodies) at a time.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.