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View additional product information for iBind™ Flex Western Starter Kit - FAQs (SLF2000S)
44 product FAQs found
以下是可能原因和解决方案：
- 转印效果差或转印不完全： 重复转印。
- 膜的转印垫不干净或污染： 使用转印垫前，用去污剂浸泡，然后用纯水彻底清洗。转印垫破损或变色后，则更换新的转印垫。
- 膜未完全湿润： 遵循预润湿膜的说明。
- 膜被指纹或角蛋白污染： 始终佩戴干净的手套并使用镊子来处理膜。处理膜时，仅触碰膜的边缘。
- 封闭不均匀： 孵育皿应足够小，使封闭液能够完全覆盖膜。
- 使用墨水标记膜： 使用墨水对膜进行标记，仅限于印迹膜的低分子量区域。
- iBind Flex卡片损坏： 更换新卡片。应确保膜在卡片上的移动仅限于标有“膜”的区域内。
- 膜与iBind Flex卡片接触不良： 加入1 mL 1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液后，立即将膜放到iBind Flex卡片上。使用附带的滚轮，确保接触良好。
以下是可能原因和解决方案：
- iBind Flex卡片损坏： 更换新卡片。应确保膜在卡片上的移动仅限于标有“膜”的区域内。
- 膜组在运行前是湿的： 应确保将5 mL 1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液加到iBind Flex卡片的平坦区。不要将溶液加到膜组上。
- iBind Flex溶液/iBind Flex荧光检测（FD）溶液准备不当： 按照使用手册（https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf）的说明，准备1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液。
以下是可能原因和解决方案：
- 蛋白质上样量过高： 减少上样量或稀释样品浓度。
- 转印效果差或转印不完全： 重复转印。
- 一抗浓度过高： 按照生产商的建议稀释一抗或通过点印迹法确定最佳抗体浓度。
以下是可能原因和解决方案：
- 转印效果差或转印不完全： 重复转印。转印后，对膜进行染色以衡量转印效率。使用阳性对照和/或分子量标记物。
- 膜未完全湿润： 遵循预润湿膜的说明。
- 一抗浓度过低： 按照生产商的建议稀释一抗或通过点印迹法确定最佳抗体浓度。
- 一抗失活： 通过点印迹法确定抗体活性。
- 一抗与抗原的亲和力较低： 获取更高亲和力的一抗。
- 二抗溶液污染： 始终佩戴手套，瓶子在未使用时始终保持拧紧状态。制备试剂时，只使用纯水。
- 目标蛋白跑出凝胶： 应确保凝胶分离范围与待转印蛋白的大小是匹配的。
- 蛋白质保留较差： 应确保凝胶分离范围与待转印蛋白的大小是匹配的。使用具有相关大小蛋白质的分子量标记物。较大的蛋白质需要较长的转印时间，较小的蛋白质所需转印时间较短。使用具有适当结合能力的膜。
- iBind Flex溶液/iBind Flex荧光检测（FD）溶液准备不当： 按照使用手册的说明，准备1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液。
- 向iBind Flex孔中加入溶液时出错： 应确保将溶液加到正确的孔中，并按正确顺序向孔中加液。
- 印迹膜在iBind Flex卡片上的位置不当： 应确保印迹膜上有蛋白质的一面与iBind Flex卡片接触，并放置在标有“膜”的区域。
- 膜组在运行前是湿的： 应确保将5 mL 1X iBind Flex溶液/iBind荧光检测（FD）溶液加到iBind Flex卡片的平坦区。不要将溶液加到膜组上。
- 不同孔中的溶液发生交叉污染： 运行期间，不要移动iBind Flex蛋白印迹处理仪。
- iBind Flex卡片损坏： 更换新卡片。应确保膜在卡片上的移动仅限于标有“膜”的区域内。
- 膜与iBind Flex卡片接触不良： 加入1 mL 1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液后，立即将膜放到iBind Flex卡片上。使用附带的滚轮，确保接触良好。
- 在运行结束前打开设备： 卡片在设备中时，不要打开设备。重新密封卡片上的孔，可导致泄漏。
- 样品制备不当；抗原性减弱或损毁： SDS和还原剂可能干扰一些抗体/抗原亲和力
- 样品太稀： 提高蛋白质样品的浓度，或增加上样量。
- 蛋白质与膜的结合较弱： 转膜液应含10–20%甲醇。
- 曝光时间不足： 对膜进行二次曝光，并延长曝光时间。
- 底物孵育不充分： 应确保每一步都达到指定时间，或在达到可接受的信噪比时，从底物中取出印迹膜。
- 底物污染： 始终佩戴手套，瓶子在未使用时始终保持拧紧状态。制备试剂时，只使用纯水。
- 印迹太旧： 蛋白质会随时间降解。应使用新制备的印迹。
以下是可能原因和解决方案：
- 膜被指纹或角蛋白污染： 始终佩戴干净的手套并使用镊子来处理膜。处理膜时，仅触碰膜的边缘。
- 一抗浓度较高： 按照生产商的建议稀释一抗或通过点印迹法确定最佳抗体浓度。
- SDS残留或转印后蛋白质与膜的结合较弱： 根据使用手册（https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf）的指示，遵循免疫检测前的膜准备说明。
- 一抗对蛋白标准品具有亲和力： 向蛋白标准品生产商咨询蛋白标准品与一抗的同源性。
- iBind Flex溶液/iBind Flex荧光检测（FD）溶液准备不当： 按照使用手册（https://tools.thermofisher.com/content/sfs/manuals/ibind_man.pdf）的说明，准备1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液。
以下是可能原因和解决方案：
- 膜未完全湿润： 遵循预润湿膜的说明。孵育皿应足够小，使封闭液能够完全覆盖膜，防止膜干燥。注意：如果使用的是PVDF膜，我们建议确保使用甲醇活化，特别是干燥的膜。刚刚结束转印并转移到1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液中的PVDF膜无需活化。
- 膜污染： 仅使用干净的新膜。始终佩戴干净的手套并使用镊子来处理膜。
- 膜过度曝光或在曝光过程中变湿： 缩短曝光时间或使信号进一步衰减。将膜放在透明薄膜中防止泄漏，并在曝光前从边缘吸去过量的底物。
- 溶液或孵育托盘污染： 使用干净的玻璃器皿和纯水制备溶液。使用新的托盘或使用玻璃器皿专用去污剂彻底清洗托盘。用纯水彻底清洗。始终佩戴干净的手套。
- 一抗浓度较高： 按照生产商的建议稀释一抗或通过点印迹法确定最佳抗体浓度。
- 对PVDF膜使用了错误的化学发光底物： 应确保使用无增强剂的CDP-Star试剂。
- 印迹膜显色过度： 遵循推荐的显色时间，当达到可接受的信噪比时，从底物中取出印迹膜。
- 使用墨水标记膜： 使用墨水对膜进行标记，仅限于印迹膜的低分子量区域。
- iBind Flex溶液/iBind Flex荧光检测（FD）溶液准备不当： 按照使用手册（https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf）的说明，准备1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液。
- 向iBind Flex孔中加入溶液时出错： 根据使用手册（https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf）的说明，按正确顺序向每孔中加入合适的溶液。
- 印迹膜在iBind Flex卡片上的位置不当：1）将膜置于iBind Flex卡片上的指定膜区域。2）印迹膜有蛋白质的一面应与iBind Flex卡片接触。3）低分子量区域应与膜组最近。-膜不应该与膜组接触。
- 卡片膜组在运行前是湿的： 应确保将10 mL 1X iBind Flex溶液/iBind Flex荧光检测（FD）溶液加到卡片的流动区域。不要将溶液加到膜组上。
以下是关于降低背景的一些其他技巧：
•如果使用iBlot PVDF膜组，应确认其尚未超过有效期，因为背景会随时间升高。使用iBlot 2 PVDF膜组将有助于降低背景。
•应确保在加入底物前将印迹膜置于蒸馏水中清洗。不要在PBS或TBS中清洗。
容易。该设备具有直观的磁性设计，用户更换插孔非常简单。但是，我们强烈建议用户在更换或洗涤时，小心处理插孔（按插槽上的指示操作），其他两个插孔在不使用时应保存在底部的托盘中。
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。
不建议这样使用，从顶部到底部不均匀流动产生的电势，可能导致穿过水平剪切膜条产生梯度信号强度。
适用，可转印单张小型印迹膜或最多3块垂直剪切膜条。但是，当使用小型或多膜条插槽时，未与卡片接触的孔应为空的。
可以。无论是否使用小型或多膜条插槽，空孔都应与卡片保持接触，应用水填充以确保所有溶液流过卡片。
当使用iBind Flex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_flex_man.pdf）第14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 Flex溶液/ iBind Flex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 Flex溶液/ iBind Flex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 Flex插槽，在下一次使用前将插槽晾干。
•小心处理插槽。
•将未使用的插槽保存于iBind Flex蛋白质印迹设备的抽屉中。
•为延长iBind Flex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_flex_man.pdf)第28页所示。
将膜置于iBind Flex溶液/ iBind Flex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 Flex溶液/ iBind Flex荧光检测（FD）溶液中额外受到封闭。
我们不建议使用您自己的溶液。iBind Flex溶液和iBind Flex荧光检测（FD）溶液具有特殊的粘稠度，并且针对iBind Flex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 Flex蛋白质免疫印迹系统（货号SLF2000）由以下组件构成：
•iBind Flex蛋白质印迹设备
•iBind Flex中型插槽
•iBind Flex小型插槽
•iBind Flex多膜条插槽
•iBind Flex印迹滚筒
下列物品需单独购买：
•iBind Flex卡片（货号SLF2010）
•iBind Flex溶液试剂盒（货号SLF2020），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 Flex荧光检测溶液试剂盒（货号SLF2019），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）。Yes. When using a conjugated primary antibody and no secondary antibody, add the diluted primary antibody to well 1 and replace diluted secondary antibody (well 3) with 1X iBind Flex/iBind Flex FD Solution. Alternatively, for faster time to results, add the diluted primary antibody to well 1, leave wells 2 and 3 empty, and add 1X iBind Flex/ iBind Flex FD Solution to well 4. This procedure will save time (~60 min) but may result in higher background.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
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.
No, we do not provide service contracts or extended warranties for these devices. The warranty is limited to 1 year from the date of shipment.
Here are possible causes and solutions:
- Poor or incomplete transfer: Repeat blot.
- Membrane pads are dirty or contaminated: Soak with detergent and rinse thoroughly with purified water before use. Replace pads when they become worn or discolored.
- Membrane not completely wet: Follow instructions for prewetting the membrane.
- Membrane is contaminated by fingerprints or keratin protein: Wear clean gloves at all times and use forceps when handling membranes. Always handle membranes around the edges.
- Uneven blocking: The incubation dish must be small enough to allow thorough coverage of membrane.
- Ink used to label membrane: Any labeling of the membrane with ink should be limited to the low MW region of the blot.
- iBind Flex Card damaged: Replace with new card. Ensure that rolling of the membrane on the card is limited to membrane region.
- Membrane is not in proper contact with the iBind Flex Card: Place the membrane on the iBind Flex Card immediately after adding a 1 mL pool of 1X iBind Flex Solution/ iBind Flex FD Solution. Use the roller provided to ensure proper contact.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Here are possible causes and solutions:
- iBind Flex Card damaged: Replace with new card. Ensure that rolling of the membrane on the card is limited to the area labeled “membrane”.
- Stack wet prior to run: Ensure that 5 mL of 1X iBind Flex Solution/ iBind Flex FD Solution is added to the flat region of the iBind Flex Card. Avoid adding solution to the Stack.
- Improper preparation of iBind Flex Solution/ iBind Flex FD Solution: Prepare 1X iBind Flex Solution/ iBind Flex FD Solution as directed in the manual.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Here are possible causes and solutions:
- Protein is overloaded: Reduce load or dilute concentration of sample.
- Poor or incomplete transfer: Repeat blot.
- Primary antibody is too concentrated: Follow the supplier's recommended dilution or determine the optimum concentration by dot blotting.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Here are possible causes and solutions:
- Poor or incomplete transfer: Repeat blot. After blotting, stain membrane to measure transfer efficiency. Use positive control and/or molecular weight marker.
- Membrane not completely wet: Follow instructions for prewetting the membrane.
- Primary antibody concentration too low: Follow the supplier's recommended dilution or determine the optimum concentration by dot blotting.
- Inactive primary antibody: Determine activity by performing a dot-blot.
- Low affinity of primary antibody to antigen: Obtain a higher affinity primary antibody.
- Contaminated secondary antibody solution: Wear gloves at all times and keep bottles tightly capped when not in use. Use only purified water when preparing reagents.
- Protein of interest ran off the gel: Match gel separation range to size of protein being transferred.
- Poor retention of proteins: Match gel separation range to size of protein being transferred. Use a molecular weight marker with relevant size proteins. Larger proteins require more transfer time, smaller proteins less. Use membrane with the appropriate binding capacity.
- Improper preparation of iBind Flex Solution/ iBind Flex FD Solution: Prepare 1X iBind Flex Solution/ iBind Flex FD Solution as directed in the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf).
- Improper application of solutions to iBind Flex Wells: Ensure that the solutions are added to the correct wells and that the wells are loaded in numerical order.
- Blot improperly placed on iBind Flex Card: Ensure that the protein side of the blot is in contact with the iBind Flex Card and is placed in the region labeled “membrane”.
- Stack wet prior to run: Ensure that 5 mL of 1X iBind Flex Solution/ iBind Flex FD Solution is added to the flat region of the iBind Flex Card. Avoid adding solution to the Stack.
- Cross-contamination of solutions in wells: Do not move the iBind Flex Western Device during the run.
- iBind Flex Card damaged: Replace with new card. Ensure that rolling of the membrane on the card is limited to the area labeled “membrane”.
- Membrane is not in proper contact with the iBind Flex Card: Place the membrane on the iBind Flex Card immediately after adding a 1 mL pool of 1X iBind Flex Solution/ iBind Flex FD Solution. Use the roller provided to ensure proper contact.
- Device opened prior to completion of run: The device should not be opened once the card has been placed in the device. Re-sealing of the wells on the card can result in leaks.
- Sample improperly prepared; antigenicity weakened or destroyed: SDS and reducing agents may interfere with some antibody/antigen affinities.
- Sample too dilute: Load a higher concentration or amount of protein onto the gel.
- Protein weakly bound to membrane: Ensure that transfer buffer contains 10-20% methanol.
- Insufficient exposure time: Re-expose film for a longer period of time.
- Insufficient substrate incubation: Perform each step for the specified amount of time or remove blot from substrate when signal-to-noise ratio is acceptable.
- Substrate is contaminated: Wear gloves at all times and keep bottles tightly capped when not in use.
- Blots are too old: Protein may have broken down over time. Use freshly prepared blots.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Here are possible causes and solutions:
- Membrane contaminated by fingerprints or keratin proteins: Wear clean gloves at all times and use forceps when handling membranes. Always handle membranes around the edges.
- Primary antibody too concentrated: Follow the supplier's recommended dilution or determine the optimum concentration by dot blotting.
- Insufficient removal of SDS/weakly bound proteins from membrane after blotting: Follow instructions for membrane preparation before immunodetection, as directed in the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf).
- Affinity of the primary antibody for the protein standards: Check with protein standard manufacturer for homologies with primary antibody.
- Improper preparation of iBind Flex Solution/ iBind Flex FD Solution: Prepare 1X iBind Flex Solution/ iBind Flex FD Solution as directed in the manual.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Here are possible causes and solutions:
- Membrane not completely wet: Follow instructions for prewetting the membrane. Use an incubation dish which is small enough to allow thorough coverage of membrane to prevent drying out. Note: If PVDF membrane is being used, we recommend making sure that it is activated with methanol, especially if it has dried up. It is not necessary to activate a PVDF membrane that has just come out of the transfer and moved into 1X iBind Flex Solution/ iBind Flex FD Solution.
- Membrane is contaminated: Use only clean, new membranes. Wear clean gloves at all times and use forceps when handling membranes.
- Film overexposed or became wet during exposure: Decrease exposure time or allow signal to further decay. Prevent leakage by encasing membrane in transparency film and blotting excess substrate from edges before exposure.
- Solutions or incubation tray are contaminated: Use clean glassware and purified water to prepare solutions. Replace or clean the tray thoroughly with a glassware-cleaning detergent. Rinse thoroughly with purified water. Wear clean gloves at all times.
- Concentrated primary antibody used: Follow the supplier's recommended dilution or determine the optimum concentration by dot blotting.
- Incorrect chemiluminescent substrate used for PVDF: Make sure CDP-Star reagent without enhancer is used.
- Blot is overdeveloped: Follow recommended developing time or remove blot from substrate when signal-to-noise ratio is acceptable.
- Ink used to label membrane: Any labeling of the membrane with ink should be limited to the low MW region of the blot.
- Improper preparation of iBind Flex Solution/ iBind Flex FD Solution: Prepare 1X iBind Flex Solution/ iBind Flex FD Solution as directed in the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf).
- Improper application of solutions to iBind Flex Wells: Add the appropriate solutions for each well in the correct numerical order as specified in the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf).
- Blot improperly placed on iBind Flex Card: 1) Place the membrane in the designated Membrane Region on the iBind Flex Card. 2) The protein side of the blot should be in contact with the iBind Flex Card. 3) The low MW regions should be closest to the Stack. 4) The membrane should not be in contact with the Stack.
- Card stack wet prior to run: Ensure that 10 mL of 1X iBind Flex/iBind Flex FD Solution is added to the flow region of the card. Avoid adding the solution to the stack.
Here are some additional tips to reduce background:
- If iBlot PVDF stacks are used, check that they have not expired as background increases with age. Using the iBlot 2 PVDF stacks will help in reducing the background.
- Make sure that the blot is rinsed in distilled water prior to adding the substrate. Do not rinse in PBS or TBS.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Yes. Due to an intuitive magnetic design, it is very simple for the user to change the well inserts. However, we highly recommend the user handle the well inserts with care (as engraved on each insert) when changing or washing, and store the other two well inserts in the convenient underside tray when not in use.
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.
It is not recommended due to the potential for non-uniform flow from top to bottom which could result in a gradient signal intensity across horizontally cut strip blots.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Yes, to run a single mini blot or up to three vertically cut strip blots. However, when using the Mini or Multi-Strip Inserts, wells not in contact with the card should remain empty.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Yes. Whether using the Mini or Multi-Strip Inserts, empty wells are still in contact with the card and should be filled with water to ensure proper flow of all solutions across the card.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Please use the protocol mentioned on Page 15 of the manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/ibind_flex_man.pdf) when using the iBind Flex 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 Flex Solution/iBind Flex 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 Flex Solution/iBind Flex Fluorescent Detection (FD) Solution.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
- Rinse the iBind Flex well inserts under running water after each use and allow the well inserts to dry before additional usage.
- Handle well inserts with care.
- Store unused well inserts in the drawer in the iBind Flex Western Device.
- To maximize the life of the springs in iBind Flex Western Device, store the device with latch unlocked, and the lid open as shown on Page 28 of the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_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 Flex Solution/ iBind Flex 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 Flex 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.
We do not recommend using your own solutions. The iBind Flex Solution and iBind Flex Fluorescent Detection (FD) Solution have specific viscosity and are optimized for the sequential lateral flow that is the principle of the iBind Flex 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 iBind Flex Western System (Cat. No. SLF2000) consists of:
- iBind Flex Western Device
- iBind Flex Midi Insert
- iBind Flex Mini Insert
- iBind Flex Multi-Strip Insert
- iBind Flex Blotting Roller
You would need to purchase the following items separately:
- iBind Flex Cards (Cat. No. SLF2010)
- iBind Flex Solution Kit (Cat. No. SLF2020) for preparing blocking, dilution, and washing buffers for chemiluminescent or chromogenic detection
- iBind Flex Fluorescent Detection Solution Kit (Cat. No. SLF2019) 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.