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Summary of Contents for SLS Blot Washer
- Page 1 Operator’s Manual For Automated Western Blot Processing...
- Page 2 Notice The information contained in this document is subject to change without notice. This document contains proprietary information which is protected by copyright. All rights are reserved. No part of this document may be photocopied, reproduced, or translated to another language without prior written consent of Stovall Life Science, Inc.
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Page 3: Table Of Contents
Inserting Tubing Into the Solenoid Valves . 9 Checking for Leaks ....... 10 Using a Second Delivery Container ..10 Cleaning the Blot Washer After Use ..12 Cleaning Tubing and Containers ... 12 Discarding Waste and Replacing Wash Buffer ........... 13 Cleaning the Exterior Surface .... -
Page 5: Chapter 1: System Familiarization And Assembly
Placement in the Laboratory Place the instrument away from sinks or other sources of water that pose a shock hazard. Do not locate the Blot Washer where the power cord will be walked on or exposed to water or chemical spills. -
Page 6: Current Requirements
Figure 1 shows the location of the fuse in a small fuse drawer on the back panel of the Blot Washer. A spare fuse is also located in this drawer. Should the fuse blow, replace with a new fuse of the same value. -
Page 7: Connecting Power
Power Receptacle Vacuum Inlet Port Fuse Drawer Figure 1. Blot washer back panel showing location of the ON/OFF power switch, power receptacle, fuse drawer, and vacuum port. Connecting Power Connect the power cord to the power receptacle on the Blot Washer back panel and connect the power cord plug to a wall or bench outlet. - Page 8 Valve #2 Lights Signaling Valve Open Figure 2. Major components on the Blot Washer front panel. Software wash programs operate three solenoid valves to control the flow of liquids. The normal condition of all valves is closed, where by the flow of liquids is impeded by pinching the silicone tubing closed.
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Page 9: Software Familiarization
During operation, the internal pump pressurizes the Wash Bottle (Figure 3) through an air pressure line. When a new wash cycle starts, Dispense Solenoid Valve #1 opens and the pressure in the Wash Bottle dispenses wash buffer from the Wash Bottle to the blot container. -
Page 10: Tube Routing
Washer, and a single blot container connected to the Blot Washer using the single dispensing harness. Color Coding Connectors on the wash bottle, waste bottle, and Blot Washer are all color coded to facilitate making the correct connections. Connecting each end of a tube to the same color connector will lead to correct connections in most cases. - Page 11 Air Pressure Tubing Wash Delivery Tubing (Blue Luer Lock) (Green Luer Lock) Blot Washer Console Wash Bottle Blue Waste Tubing Black Delivery Vacuum Tubing Tubing (Black Luer Lock) Waste Tubing (Orange Luer Lock) Western Blot Incubation Box Waste or Similar Blot...
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Page 12: Connecting The Waste Removal System
For best performance, place the Wash Bottle above the blot container. Begin by connecting air delivery tubing from the blue luer lock on the left side of the Blot Washer front panel (Figure 2) to the blue luer lock connector on top of the Wash Bottle. Next... -
Page 13: Inserting Tubing Into The Solenoid Valves
delivery system is completed by connecting tubing from the single dispensing harness to the right side connector on the manifold and to the delivery bit in the blot container. The bottom of the delivery bit should touch the bottom of the blot container. Inserting Tubing Into the Solenoid Valves The solenoid valves are closed until opened by manual or program command. -
Page 14: Checking For Leaks
Notice that the air pressure provided by the Blot Washer’s pump is divided with a “Y” connector in the tubing. The wash delivery tube from the Wash Bottle is routed through Dispense Valve #1, as... - Page 15 140 ml syringe or second delivery container Figure 5. Tubing configuration for using the 140 ml syringe or other secondary delivery container (waste tubing not shown).
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Page 16: Cleaning The Blot Washer After Use
Cleaning of reagent containers and tubing is the primary mainte- nance requirement. To eliminate almost all potential maintenance problems, establish the habit of cleaning the Blot Washer immedi- ately after use. Reagent residues are easy to clean up while they are fresh and still wet. -
Page 17: Discarding Waste And Replacing Wash Buffer
Blot Washer. Warning: Do not immerse the Blot Washer in water or other liquid. The exterior of the Blot Washer can be cleaned with a rag dampened with a solution of mild detergent. Rinse by wiping with a cloth dampened in water. -
Page 19: Chapter 2: Creating And Running Programs
– Toggles through available options for a step. – Accepts the current option and moves to the next step. Powering On the Blot Washer 1) Turn on the power switch on the back of the Blot Washer control console. The initial display is used to select SELECT NUMBER OF the number of blots. -
Page 20: Selecting The Program Number To Edit Or Run
16 CHAPTER 2 Creating Wash Sequences “Single” is always the correct choice unless the Quad Harness is being used. See Using the Quad Harness below to learn how to enter programs for washing 1 to 4 blots simultaneously. Selecting the Program Number to Edit or Run 2) Press to accept “Single”... - Page 21 Each program can have up to four steps and each program step can be one of three types: DELAY, WASH, or INCUBATE. DELAY A time delay during which no actions are performed. A typical use is to add DELAY as the first step to allow for incubation of previously administered primary antibodies.
- Page 22 18 CHAPTER 2 Creating Wash Sequences Any of the three types of steps can be assigned to a given step. Examples of typical programs are given below. # Steps Typical Steps DELAY, WASH, INCUBATE, WASH WASH, INCUBATE, WASH WASH, WASH (two different wash buffers from different containers) WASH In our continuing example, program #1 will be edited to be a...
- Page 23 WASH VALVE: 1 Select WASH VALVE 1 OR 2 to dispense wash buffer from Dispense Valve #1 or Dispense Valve #2 on the Blot Washer front panel (see Figure 2, Chapter 1). Typically Dispense Valve #1 is used unless you want to dispense from a secondary delivery container.
- Page 24 INCB VALVE: 2 Typically, secondary antibodies for the incubation step are dispensed from the accessory syringe or a second reservoir bottle using Dispense Valve #2 on the Blot Washer front panel. This configuration will be assumed for this example program. 14) Press to accept Dispense Value #2.
- Page 25 Note: The dispense volume entered should not exceed the volume of the 140 ml accessory syringe or other secondary delivery con- tainer. 15) Press to accept the DISPENSE volume. EDIT P1 STEP 3 INCB TIME: 01:00 Set the incubation time (HH:MM) according to your protocol by pressing the keys.
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Page 26: Starting A Program
22 CHAPTER 2 Creating Wash Sequences Starting a Program 1) Turn on the Blot Washer, if necessary. The initial display is used to select SELECT NUMBER OF the number of blots. BLOTS: SINGLE 2) Press to check the system pressure. -
Page 27: Pausing A Program
Pausing a Program While a program is in operation it can be paused by pressing The program number and current RUN P 1 STEP 1 step are listed in the top line of the display. The second line indicates PAUSED operation is paused. - Page 28 Quad Harness to the right side of the manifold on the Blot Washer front panel. 2) Turn on the Blot Washer and wait until prompted for the number of blots. The initial display is used to select SELECT NUMBER OF the number of blots.
- Page 29 Figure 7. Quad har- ness connected to four Western Incu- bation Boxes on a shaker (The Belly Dancer, Stovall Life Science, Inc., Greensboro, NC). 4) Press to set the number of blots and continue. SELECT PROGRAM NUMBER: 1 5) Press once to increment the program number.
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Page 30: Scaling Up Or Back
40 ml of wash buffer, the appropriate WASH step in the program should be set to dispense 40 ml. Blot Washer already knows that 40 ml should be delivered to each container if 2 OF QUAD, 3 OF QUAD, or 4 OF QUAD is chosen for NUMBER OF BLOTS. - Page 31 Program # Description 1-10 4-step DELAY, WASH, INCUBATE, and WASH pro- grams. 11-13 3-step WASH, INCUBATE, and WASH programs. 14-16 2-step WASH, WASH programs. 17-19 1-step WASH programs. A 4-step test program used during factory testing that can be modified in the same way as the other 19 pro- grams.
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Page 33: Chapter 3: Calibration
Chapter 3: Calibration The dispense rate and aspiration rate are factory calibrated. Volume of liquids delivered or aspirated is determined by the length of time the solenoid valves are open. For delivery, the pump maintains 23 KPA of pressure, which results in a dispense rate of approximately 2.5 ml/sec;... -
Page 34: Checking System Pressure
CHAPTER 3 Calibration Checking System Pressure One factor that affects the dispense rate is the maintenance of air pressure in the liquid containers. System pressure should always be checked before proceeding to other calibration steps. 1) Press at the initial SELECT NUMBER OF BLOTS display to check the system pressure. -
Page 35: Setting The Aspiration Vacuum Factor
7) Hold the 100 ml graduated cylinder while it fills at a level as close as possible to the level of the membrane boxes when operating the Blot Washer for automating Western Blot processing. Setting the Aspiration Vacuum Factor... -
Page 37: Accessory Part Numbers
Chapter 4: Appendix Accessory Part Numbers ACCBW0002 4L Wash Bottle ACCBW0003 140 ml Lock Syringe ACCBW0005 Quad Pumping Harness ACCBW0001 Collet w/Delivery Tube Three different sizes of Stovall Western Incubation Boxes are available in various pack sizes. See www.slscience.com. Specifications Console Dimensions: 23 W x 9.6 D x 5.4 cm H (9”... - Page 38 34 CHAPTER 4 Appendix Program Storage Capacity: 20 programs with up to 4 steps per program. Liquid Flow Control Valves: 2 for delivery, 1 for aspiration.
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Page 39: Declaration Of Conformity
Stovall Life Science, Inc. 206 G. So. Westgate Drive Greensboro, North Carolina, 27407 USA Declares under our sole responsibility that the Blot Washer is in conformity with the following EMC Standards: EN50081-1: 1997 Generic emission standards for residential commercial, and light industry environments... - Page 40 Printed Franklin’s 30 Units 4-2-10...
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