GO-Blot™ V2: Fully Automated and Programmable Western Blot Processor
A Western blotting device has never been so flexible and easy to use as the new GO-Blot™ V2. There are three tray sizes to choose from and infinite (nearly) combinations of Protocols to create. In addition, there is a growing community of users that share their successful protocol/antibody combinations on the Global Western Portal™.
The GO-Blot™ V2 Western Blot Processor is a reliable fluid delivery device for reproducibly probing blots and membranes with primary and secondary antibodies. It was developed after communication with hundreds of research scientists over the past two years. They required a cost-effective machine that would reproduce the hands-on method using a rocker platform and exchanging buffers manually.
The equipment is composed of valves, pumps, a motorized tilting platform and a control board that coordinates timing with the application of the correct solution to the membrane or blot. It can operate at room temperature or in a cold room*. All commonly used buffers are applicable, e.g. PBST and TBST. The purchase of the machine comes with a full warranty (parts & labor or replacement) for up to one year after the date of purchase.
* - Note cold room operation on order if applicable.
The GO-Blot™ V2 allows the operator to load their own membranes and add their blocking solution, primary and secondary antibodies to reservoirs. The machine handles the rest of the western blot processing. Plus, the GO-Blot V2 makes it easy to recycle your primary antibody.
Choose from 10 Pre-defined Protocols or try out the new phone interface to create your own protocol. Visit mygoblot.com™ to set up your user account, and store your results to share with your lab or the global community.
The machine is designed to be modular and economical so that you utilize multiple machines running at different times with different routines and different antibodies, in other words, it’s very flexible for multiple users. Also being modular means that you can add new units when the budget allows or your project demands it.
Create your own routines
You can program your own runs through the easy to use mygoblot.com website through your phone and share them with users around the world using the globabl western portal. Labs which require more secure options are not required to share their runs or results.
It is important to pour and pipette accurately into the Wash, Primary and Secondary reservoirs because liquid overflows can result in equipment malfunction. To ensure good technique pour Wash buffer into the middle of the Wash Buffer reservoir, and for the syringe loading technique use a pipettor to pipette into the syringe at a depth that is below the rim.
The most generally applicable blocking solution is TBST with 5% (w/v) dried milk powder. However other blocking reagents are possible such as 1% BSA or 1% non-specific IgG. TBST is Tris buffered saline with Tween 20 which contains 10 mM Tris-HCl pH 7.0, 150 mM NaCl and 0.05% (v/v) Tween 20.
The best volume of primary antibody is 10 ml, at this volume all routines run efficiently and will not be subject to the effects of evaporation.
Dilute primary antibody in TBST or if this concentration produces high background then use TBST with 1% milk.
The GO-Blot™ V2 can be operated in a cold room at 4°C, but it is not recommended to move the machine to room temperature in the middle of a routine because if the power is disconnected the program will need to be re-started from the beginning, hence routines should be run at one temperature.
The secondary antibody is raised to the primary IgG species and usually is conjugated to horse-radish peroxidase (HRP) or alkaline phosphatase (AP).Dilute secondary antibody 1:20,000 in TBST or if this concentration produces high background then use TBST with 1% milk.
The device is used by Cytoskeleton Inc. research and development scientists to characterize and validate the new line of antibodies.
US patents pending.
Do not attempt to repair the GO-Blot V2. If needed, the device should be returned to Cytoskeleton Inc. for new parts. If the warranty is still valid then the repairs will be free of charge, otherwise a Purchase Order or credit card is required for the repair and parts fee.
Creating a Protocol
Open a browser in a computer or phone.
Type in the IP address of the GO-Blot V2 into the address bar and tap Enter or Go respectively.
1. Tap on “Create Protocol”
2. Fill out each field with appropriate data, (e.g. the wash steps have two data points, the first one is number of washes, the second is length of each wash in minutes.)
3. When it is complete, tap “Submit” to save the new Protocol.
Note: New Protocols are listed chronologically, therefore the latest protocol will be at the end of the list.
Note: The Large tray type will be limited automatically if there are too many washes, the maximum number of washes for the Large tray Is 12 total.
Tip: If you cannot see the Submit button on the iphone version, shrink the screen and/or tap on AA symbol in the address bar, and chose “Hide address bar” option.
Tip: Pause buttons are useful to pause the run while adding a labile reagent or checking a routine’s timing.
Running a Protocol
Warning: do not load solutions until the Water Wash Protocol has run.
A Water Wash is necessary to start using your GO-Blot V2 and to ensure all systems are working well. A Water Wash is also recommended after greater than 1 week without a run. Sometimes the last operator might forget to clean the machine with a Water Wash, in this situation the Water Wash must be run before using the machine.
1. While standing next to the GO-Blot V2 check that no solutions are in any reservoirs or trays.
2. Connect the luer locks with tubing to the front of the machine.
3. Place the left and right-side tubing into a waste container.
4. While on the Main Menu, scroll down by clicking the machine’s “Down” button until you reach the Run Protocols line.
5. Click the “Enter” button.
6. Scroll down to highlight the Water Wash, and click Enter.
7. An alert should pop-up indicating to fill the Water Wash reservoir, lets do this by adding 425 ml to the container which covers the level sensor.
8. The alert should disappear and the option to Run the protocol appears, click Enter to run the protocol.
Note: The same approach is used to run a protocol, except you need to add Blocking solution and membrane to the tray, TBST (or PBST) to the Buffer reservoir and appropriate Primary and Secondary antibody solutions to their respective receptacles. Also, place the right-side “Recycle” tubing into a clean recycle vessel (e.g. 15 or 50 ml disposable tube). Then chose a protocol from the list. Pre-defined Protocols are at the top of the list and User defined (with a “U” before the number) ones at the end of the list.
Warning: Be sure to visit the machine and view the screen before adding antibody and buffer solutions. If a previous operator did not run the Water Wash, then it will be required before continuing, an alert will be visible, any buffers and antibody solutions loaded at this point will be washed away!
Note: If there is a sticky-high affinity antibody left over from a previous run. The best way to clean the device is to add 50 ml of either 1 x SDS-running buffer from a PAGE system (e.g. a standard composition is 50 mM Tris-HCl pH 6.8, 100 mM NaCl, 0.1% SDS) to all the reservoirs. Then choose the Water Wash protocol to run the solution through the system. Then follow with two Water Washes using pure water only.
User and password
The original user will receive the login credentials by email. please contact firstname.lastname@example.org if you do not have access to it.
1. Logon to https://mygoblot.com
2. Click on the fourth bar down called, “Personal Western Archive”
3. A list will pop-up.
4. Click on the one you want to open
Tip – You can select the name or date range by the table’s title drop down menus.
Uploading results to complete an archive
Note: The empty boxes are ones you can fill in with experimental data and save, or save and submit for publication on the Global Western Portal.
1. Click in a box to start adding text.
2. Click on a yellow + sign to add a hyperlink
3. Click on upload files to add your experiment-tal blot image.
4. Click Save to save data.
Note: Partially filled out fields are fine to save, but empty field files cannot be submitted to the Global Western Portal.
Submitting a Routine to the Global Western Portal
1. Fill out all fields of a Personal Western Archive.
2. Click Save to save content.
3. Scroll to the end of the file and locate the “Submit for Approval” button, and click it.
4. With two days you will receive a notification of approval or additional questions before approval.
Downloading a Routine from Global Western Portal
1. Go to the homepage of https://mygoblot.com
2. Click on Global Western Portal.
3. Using the table’s title search field, e.g. type in the Antigen of interest.
4. A short list of matches will pop-up. Click on one row.
5. Scroll to the bottom of the record, and click the “Share/Download” button.
6. Chose one of the options and click on it, a “json” file will download.
7. Receiving on the phone requires opening the email app and “sending” the json file to the Downloads folder.
Importing a file to a GO-Blot V2 device.
1. Open the GO-Blot V2 device control panel in a browser (see Section 4 above).
2. Click Create Protocol.
3. Scroll down to the “Import file” button.
4. Select the file from your download folder.
Tip: If you cannot see the Submit button on the phone version, shrink the screen and/or flick the address bar off the top of the screen e.g. on iPhone tap on AA symbol at top left, and chose “hide address bar” option.
Tip: Download folder locations vary, on the iPhone they are located in the Files App > Downloads folder, whereas for the Android they are stored in My Files or File Manager > Downloads. On the PC computer they are found in the main C:/Download folder, or for Apple computers in the Users>Downloads folder or the Finder sidebar.
Setting up users in mygoblot.com
Note: When you purchase a new GO-Blot V2 your name will be used as the main user (i.e. administrator), yes you have become an administrator! Also, you will be given a Serial Number, keep both of these close to the GO-Blot V2 so we can identify it if needed later.
1. Login to https://mygoblot.com using your user name and Serial Number.
2. Go to the My GO-Blot Devices button and you will see your machine. if you do not see it call Customer Support (303-322-2254 or email email@example.com .
3. Now go back to the home page by clicking on top of the webpage, and click the “Register User” button.
4. Type in the User’s credentials and click Save.
Note: The person who bought the GO-Blot V2 unit will be the Main User, all others are called Lab Users. If this needs to change contact Customer Service at the email above.
Note: For multiple GO-Blots it is useful to have the same person buying them so that the Main User stays the same and all other lab personnel are entered via the Main User.
Setting up a direct connection from phone to GO-Blot V2
This connection method is the most simple and direct, it allows the user to quickly access the programming and protocol tracking functions from a mobile phone.
1. Click the “Down” button, to Connectivity.
2. Click “Enter” button.
3. Scan the QR code.
4. Choose GO-Blot V2 Wifi connection from your phone’s Settings>Wifi location. Make sure this is connected by seeing the check (tick) mark.
5. Go to the phone’s browser, and type in 10.1.1.1 and hit Enter or GO.
6. A green page with the GO-Blot V2 menu in green will pop up.
7. Tap Create Protocol to start programming the steps for your unique Western Protocol.
Setting up a network Wifi connection to access mygoblot.com
This method allows full functionality of mygoblot.com such as offsite tracking of protocol progress, user registration, access to archive runs, and ability to add data to the webpage and submit a protocol to the Global Western Archive.
1. Starting from the direct access mode described in the previous section, go to the home page of 10.1.1.1 IP address.
2. Click on Settings and then Server Setup.
3. Type in the following:
a) your network’s Web name or Wep ID,
b) Web password,
c) a new IP address obtained from your IT department
d) the Gateway IP and Port # from your IT department.
Note1: This information will be obtained from your IT department.
Note2: Some phones have the Save button below the screen’s view, in that case either shrink the screen and/or tap on AA symbol in the address bar, and chose “Hide address bar” option.
4. When the IP is confirmed, make a label and attach to the GO-Blot V2 in an obvious permanent place. Next time anyone wants to use the device, they can just type this number into their phone’s web browser.
5. Click “Save” to complete the process.
6. Click on Date & Time button.
7. Enter your local zone details (e.g. GB and France are 00.00, New York is -05.00, San Francisco is -08.00, Eastern Australia is +10.00 and Beijing is +8.00 and Japan +09.00), then tap on month and year to change them, and click “Save”. Note: Some phones have the Save button below the screen’s view, in that case either shrink the screen and/or tap on AA symbol in the address bar, and chose “Hide address bar” option.
8. Close browser, and click “Back” on the GO-Blot V2 device.
9. Open a browser on your phone, type in the IP and see the control panel in the browser.