Biotool Protein Transfer System

Transcription

1 A Copany of Selleck Cheicals Instruction Manual Protein Transfer Syste

2 Table of Contents Safety Warning Introduction 2 Setup 3 Biotool Protein Transfer Stack 4 Protein Transfer Flow Chart 5 Protein Transfer Protocol 6 Consuables and Related Products 9 Trouble Shooting 0 Safety Warning! DO NOT open or attept to repair the Biotool Protein Transfer Syste or Cassettes. Doing so will void your warranties and can put you at risk for electric shock. Please contact Biotool and return or Cassettes to Biotool if repairs are needed. Under noral operating conditions, Biotool Protein Transfer Syste ay becoe war during operations.

3 Introduction The utilizes an optiized, accelerated, and highthroughput sei-dry protein transfer protocol. Using this syste and the readyto-use Biotool Protein Transfer Stack, the protein transfer can be copleted as quickly as 0 inutes with excellent transfer efficiency and reproducibility. In addition, with two pre-prograed and independently-running Biotool Transfer Cassettes, the transfer of 4 ini-forat gels ( c) or 2 idi-forat gels ( c) can be processed at the sae tie. Contains: Biotool Protein Transfer Instruent Biotool Transfer Cassette 2 Biotool Tweezers Power Supply Cord Instruction Manual White Plate Blotting Box Gel Cutter 2

4 Setup. Place the syste on a level surface with at least 0 c of clearance in the back for proper ventilation to ensure adequate cooling. 2. Plug the power supply cord into the back of the instruent, and connect it to a standard grounded outlet. 3. Reove the foa ring separating the instruent and cassettes. 4. Power on the instruent using the switch on the right side of the unit. LCD Display Cassette Navigation Buttons Plate Lap Protein Transfer Syste USB Port Power Switch Notice: Do not place heavy ites on the top of the Biotool Protein Transfer o Syste. The should be used at 5 30 C abient teperature and 0 95% relative huidity (noncondensing). Do not operate in extree huidity (>95%) or where condensation can affect the internal electrical circuits of the unit. Always connect the unit to a grounded AC outlet using the power supply cord provided. This instruent should not be odified or altered in any way. Alteration of this instruent will void the anufacturer s warranty. Iportant: This instruent is intended for laboratory use only. 3

5 Biotool Protein Transfer Stack Contents B824 (ini, 000 ) (idi, 0 ) Top Stack Botto Stack Balance Buffer (L) (ini, 20 ) B825 (ini, 00 ) B826 B834 B835 (idi, 00 ) B836 (idi, 000 ) Notice: Store at roo teperature. For research use only. Please wear gloves and protective eyewear when handling. The Biotool Protein Transfer Stack is for single-use only. 4

6 Protein Transfer Flow Chart Mebrane Gel (NC or PVDF) ddh2o Balance Buffer ce Balan Pre-Treatent Buffer Botto 5s 30s Botto Top Top Top Stack BOTTOM Stack For video instructions,please visit: For video instructions,please visit: Sandwich Assebly Top Stack Botto Stack Cassette Setup.5 Step Ste p 2.0 Protein Transfer Syste Progra Setup Cell A: Transfer Cell A: MINI*/MINI*2/MIDI* Cassette Connected Gel setting Cell A: 5:00 Cell A: Cell B: Cell C: Cell D: Tie setting Cell A: Transfer One MINI Gel for 5:00 Start/Cancel Cell A: Copleted Progra Cofiration running... Transfer Copleted After Treatent ( See Protocol Step7) 5

7 Protein Transfer Protocol. Soak the NC or PVDF ebranes in Balance Buffer for 30s. Note: Do not pretreat PVDF ebranes using ethanol. The Balance Buffer contained in Biotool Protein Transfer Stack can activate PVDF directly. 2. Soak the SDS-PAGE in ddh2o for 5s to reove electrophoresis buffer. 3. Asseble transfer sandwich. Each Biotool Transfer Cassette can hold or 2 ini forat gels ( c) or one idi forat gel ( c). Different or identical protocols can be set up for each cassette. Top Cassette Top Stack Gel Mebrane (NC or PVDF) Botto Stack Botto Cassette () Open the Biotool Transfer Cassette and place the BOTTOM Stack close to the c of the cassette botto. Notes: Seal the package iediately after use to prevent evaporation, which ay decrease the transfer efficiency. It is not necessary to re-position an assebled transfer pack if it is only slightly off c in the cassette. 6

8 (2) Gently put a pre-soaked ebrane (NC or PVDF) (treated in the st Step) on the Botto Stack. Note: The ebrane should be cut properly to fit the size of the stack. (3) Place a ddh2o-rinsed SDS-PAGE gel (treated in 2nd Step) on the ebrane. Then, GENTLY eliinate air bubbles between gel and ebrane using the WB Roller. Note: The gel should be cut properly to fit the size of the stack. All air bubbles between the gel and ebrane MUST be eliinated. (4) Put the TOP Stack on the gel. Note: Seal the package iediately after use to prevent evaporation, which ay decrease the transfer efficiency. 4. Place the cassette lid on the base. Press the lid down and turn the dial clockwise to engage all the four lid pins into the locking slots on both sides. Then keep turning the dial clockwise until it reaches your gel thickness (.5,.0, 0.75, or 0.5 )..5.5 Step St ep Gel cassette Turn on the. Slide the cassette (with the dial facing up) into one of the bays until it akes contact with the agnetic interlock in the back of the instruent tub and you can hear a click. 7

9 6. Progra setup () Select the bay in which the Cassette is inserted (2) Choose the gel forat. Cell A: Transfer Cell A: MINI*/MINI*2/MIDI* to select to select to confir to confir (3) Set the transfer tie according to the following table: MW Gel Thickness Tie Standard ( 00 kda) in High (>00 kda) 0.5 Cell A: 5:00 5 in to adjust tie in to confir (4) Start Transfer. The tie will count down (5) The LCD enu screen will display until the transfer process is coplete. Copleted at the end of the transfer. Cell A: Cell B: Cell C: Cell D: to confir Cell A: Transfer One MINI Gel for 5:00 Start/Cancel Cell A: Copleted Note: The can be upgraded to containing 4 Cassettes for 4 independent bays. Please contact Biotool if needed. 8

10 7. Pull the cassette straight out fro the bay and unlock it by turning the dial counterclockwise to the Unlock position. Disasseble the blotting sandwich and place the ebrane iediately into the Balance Buffer for 30s to re-activate and to avoid dry-out. Then rinse it using TBST for 5s before further application. Note: Be careful when reoving a cassette fro the unit after a transfer run. Soe parts of the cassette ay be hot. Discard all the transfer stacks after use, they are not reusable. 8. Cleaning () Epty residual liquid fro the cassette. Rinse the base and lid of the cassette with deionized water and dry the with a paper towel. (2) Before cleaning, ensure the syste is not in use and then turn off the syste with the power switch. For instruent cleaning, wipe with a rag or absorbent paper. You can also use 20% ethanol if necessary. For cassette cleaning, wipe with a rag or absorbent paper. You can also use the tap water to rinse down cassette. Consuables and Related Products 9 Cat. No. Product Nae Size B824 Protein Transfer Consuables (ini, 20X) B825 Protein Transfer Consuables (ini, 00X) B826 Protein Transfer Consuables (ini, 000X) 20 RUNs 00 RUNs B834 Protein Transfer Consuables (idi, 0X) B835 Protein Transfer Consuables (idi, 00X) 0 RUNs 00 RUNs B836 Protein Transfer Consuables (idi, 000X) 000 RUNs B800 Protein Transfer Syste set B800 Gel Cassette set 000 RUNs

11 Trouble Shooting Please review the following for trouble-shooting opinions when you encounter technical difficulties. Alternatively, feel free to contact Biotool technical support (Eail: Tel: ) directly. Proble Potential Cause(s) Suggestion(s) Poor or no electrophoretic Setup error Setup the right progra (Transfer gel s Nuber) transfer; excessive Positioning error Choose a suitable gel protein reains in the gel (as detected by staining the gel after thickness (0.75/.0/.5 ) Transfer tie is not long enough. transfer) High olecular weight proteins (> 50 kda) ay require increased transfer tie Swirls or issing Poor contact between the Use the Roller to ensure that bands ebrane and the gel. Air air bubbles are reoved. diffuse transfers bubbles reain between the ebrane and gel. None is displayed The Instruent is not Check the sandwich and on the Run screen when the prepared detecting the presence of a cassette ake sure the TOP and BOTTOM Stacks are OK. cassette is in the Ensure cassette is firly Instruent inserted into the bay. Expired reagents Use the reagents within Equipent failure Contact Technical Support labeled shelf life for assistance. 0

12 For video instructions please visit Order & Inquiry Tel: Fax: E-ail: order@biotool.co