GE Amersham ECl Gel Box User Manual
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GE Amersham ECl Gel Box User Manual

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GE Healthcare
Life Sciences
Amersham™ ECL™ Gel Box
User Manual

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Summary of Contents for GE Amersham ECl Gel Box

  • Page 1 GE Healthcare Life Sciences Amersham™ ECL™ Gel Box User Manual...

  • Page 3: Table Of Contents

    Safety precautions ..................... Labels ..........................Recycling information ....................Declaration of Hazardous Substances (DoHS) ..........System description ................Amersham ECL Gel Box .................... Amersham ECL Gel ..................... Installation ..................Protocols ....................Electrophoresis protocol ..................Removing the gel from the cassette ..............

  • Page 5: Introduction

    1 Introduction Introduction General information Amersham ECL Gel Box together with Amersham ECL Gel constitute a ready-to-use mini-gel system for high quality separation of proteins. Amersham ECL Gel is compatible with Tris-Glycine buffer and contains no SDS, making it ideal for protein analysis under both native and denaturing conditions.

  • Page 6: Protein Gel Electrophoresis Overview

    1 Introduction 1.1 Protein gel electrophoresis overview Protein gel electrophoresis overview Below is a flowchart of protein analysis including separation of proteins using PAGE followed by either Western blotting or post-staining detection methods. Gel electrophoresis Western blotting Post-staining detection method detection method Transfer Post-staining...

  • Page 7: Important User Information

    Important user information Intended use Amersham ECL Gel Box and Amersham ECL Gel are intended for PAGE. Amersham ECL Gel Box and Amersham ECL Gel are intended for research use only, and shall not be used in any clinical procedures, or for diagnostic purposes.
  • Page 8 1 Introduction 1.2 Important user information Notices NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment. Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...

  • Page 9: Regulatory Information

    • used in the same state as it was delivered from GE Healthcare, except for alterations described in the user documentation. Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...
  • Page 10 Regulatory compliance of connected equipment Any equipment connected to Amersham ECL Gel Box should meet the safety requirements of EN 61010-1/IEC 61010-1, or other relevant harmonized standards. Within EU, connected equipment must be CE marked. Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...

  • Page 11: Safety

    2 Safety Safety About this chapter This chapter presents safety precautions for Amersham ECL Gel Box. Labels on the instrument and information regarding recycling are also included in this chapter. In this chapter This chapter contains the following sections: Section page 2.1 Safety precautions...

  • Page 12: Safety Precautions

    2 Safety 2.1 Safety precautions Safety precautions General precautions WARNING Do not use Amersham ECL Gel Box in any other way than described in the Amersham ECL Gel Box Instructions. CAUTION Operation of Amersham ECL Gel Box should be performed by properly trained personnel only.
  • Page 13 Gel Box is completely dry on the outside before connecting it to a power supply. Wipe dry with a cloth. WARNING Do not use Amersham ECL Gel Box if it is not working properly, nor if it has suffered any damage, for example: •...
  • Page 14 Amersham ECL Gel Box from the power supply before removing the safety lid. WARNING Do not operate Amersham ECL Gel Box without a full- functioning safety lid in place. WARNING Do not exceed the maximum operating voltage of...
  • Page 15 High voltage. Do not overfill Amersham ECL Gel Box with running buffer. Maximum volume is 90 ml per tank. WARNING Do not operate Amersham ECL Gel Box close to strong magnetic fields. CAUTION Hazardous substances. When using hazardous chemicals, take all suitable protective measures, such as wearing protective glasses and gloves resistant to the substances used.
  • Page 16 WARNING Cleaning. Do not autoclave, bake or microwave Amersham ECL Gel Box. WARNING Cleaning. Do not wash Amersham ECL Gel Box in a mechanical washer. WARNING Cleaning. Do not use abrasive creams or scourers. Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...
  • Page 17 2 Safety 2.1 Safety precautions NOTICE Cleaning. Clean the instrument with distilled water and wipe dry with a soft damp tissue. Let the instrument dry completely before use. Materials Safety Data Sheet It is recommended to read the Materials Safety Data Sheet (MSDS) before using Amersham ECL Gel.

  • Page 18: Labels

    Labels Introduction This section describes the safety labels and labels concerning hazardous substances that are attached to the Amersham ECL Gel Box instrument. Labels on Amersham ECL Gel Box The illustration below shows an example of the label that is attached to the bottom of the Amersham ECL Gel Box.
  • Page 19 2 Safety 2.2 Labels Label Meaning Warning! High Voltage. Always make sure that the instrument is disconnected from electric power before opening the lid or disconnecting any electric equipment. The instrument complies with the requirements for electromagnetic compliance (EMC) in Australia and New Zealand.
  • Page 20 2 Safety 2.2 Labels Labels concerning hazardous substances The following symbols on the labels concern hazardous substances: Label Meaning This symbol indicates that electrical and electronic equipment must not be disposed of as unsorted municipal waste and must be collected separately. Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment.

  • Page 21: Recycling Information

    Disposal, general instructions When taking Amersham ECL Gel Box out of service, the different materials must be separated and recycled according to national and local environmental regulations. Disposal of electrical...

  • Page 22: Declaration Of Hazardous Substances (Dohs)

    2 Safety 2.4 Declaration of Hazardous Substances (DoHS) Declaration of Hazardous Substances (DoHS) Introduction The following product pollution control information is provided according to SJ/T11364-2006 Marking for Control of Pollution caused by Electronic Information Products. 根据SJ/T11364-2006《电子信息产品污染控制标识要求》特提供如下有 关污染 控制方面的信息 Symbols used in pollution control label 电子信息产品污染控制标志说明...
  • Page 23 2 Safety 2.4 Declaration of Hazardous Substances (DoHS) Label Meaning Consumables or certain parts may have their own label with an EFUP value less than the product. Periodic replacement of those consumables or parts to maintain the declared EFUP shall be done in accordance with the Product Maintenance Procedures.
  • Page 24 2 Safety 2.4 Declaration of Hazardous Substances (DoHS) List of hazardous substances and their concentrations 产品中有毒有害物质或元素的名称及含量 Indication for each major part if substance exceeds limit Value Meaning Indicates that this toxic or hazardous substance contained in all of the homogeneous materials for this part is below the limit requirement in SJ/T11363-2006.
  • Page 25 2 Safety 2.4 Declaration of Hazardous Substances (DoHS) List of hazardous substances Component Hazardous substance name 有毒有害物质或元素 部件名称 Cr6+ PBDE 铅 汞 镉 六价铬 多溴联苯 多溴二苯醚 Amersham ECL Gel Box, 28-9902-52 The product has not been tested as per the Chinese standard SJ/T11363-2006 Requirements for Concentration Limits for Certain Hazardous Substances in Electronic Information Product.

  • Page 26: System Description

    3 System description System description About this chapter This chapter describes the features of Amersham ECL Gel Box and Amersham ECL Gel. In this chapter This chapter contains the following sections: Section page 3.1 Amersham ECL Gel Box 3.2 Amersham ECL Gel...

  • Page 27: Amersham Ecl Gel Box

    3.1 Amersham ECL Gel Box Amersham ECL Gel Box Introduction This section describes the features of Amersham ECL Gel Box. Overview The illustration below shows the main parts of Amersham ECL Gel Box. Part Description Part Description Power cord (to external...
  • Page 28 3 System description 3.1 Amersham ECL Gel Box Gel Box specifications Parameter Value Dimensions: Gel Box 167 x 148 x 43.5 mm (W x H x D) Gel cassette 116.8 x 84 x 15.6 mm (W x H x D)
  • Page 29 Safety lid Amersham ECL Gel Box is equipped with a safety lid, which shall be placed on top of the base unit before a power supply is connected. The safety lid has two magnets which will activate current only when their magnetic fields are in contact with the electrodes inside the base unit of Amersham ECL Gel Box.

  • Page 30: Amersham Ecl Gel

    Amersham ECL Gel Introduction This section describes the features of Amersham ECL Gel. Design Amersham ECL Gel is designed for PAGE using Amersham ECL Gel Box. Gel concentrations Amersham ECL Gels are available as homogenous and gradient gels in the following concentrations: •...
  • Page 31 3 System description 3.2 Amersham ECL Gel Separation range The following image shows the separation pattern of proteins at different molecular weight for each Amersham ECL Gel. 4-12 8-16 4-20 % Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...
  • Page 32 3 System description 3.2 Amersham ECL Gel Gel specifications Parameter Value Gel matrix Acrylamide/bis-acrylamide Gel concentration %C=2.6% (w/w) %T=Total concentration (w/v) of acrylamide + bisacrylamide is given as percentage in the respective Amersham ECL Gel product name. Gel thickness 1.4 mm Gel size 7.5 cm x 8.0 cm (W x H) Sample well...
  • Page 33 3 System description 3.2 Amersham ECL Gel Buffer solutions Solution Preparations 2× SDS Sample 1 Mix 2.5 ml of 0.5 M Tris-HCl, pH 6.8, 4.0 ml of buffer 10% (w/v) SDS, 2.0 ml of glycerol, 2.0 mg of bromphenol blue and 310 mg of DTT. 2 Make up to 10 ml with distilled water.

  • Page 34: Installation

    4 Installation Installation In this chapter This chapter describes how to connect Amersham ECL Gel Box to a power supply. Precautions WARNING Access to power switch and power cord. Do not block access to the Power switch on the Power supply and the Power cord.
  • Page 35 4 Installation Connecting GE Healthcare power supplies Connect Amersham ECL Gel Box to the power supply according to the instructions supplied with the power supply. Connecting other power supplies The power supply must have a rigid insulating sleeve output connector that fulfills the requirements for 200 V (DC), (for example EPS 301).
  • Page 36 4 Installation Power supply compatibility guide for Amersham™ ECL™ Gel Box Power supply type Adapter Example Equipped with Ø 4 Not needed GEHC EPS 301 mm rigid insulation Without Ø 4 mm rigid Adapter supplied with insulation; inner dis- Amersham ECL Gel tance >6 mm High-voltage power 18-1129-59...
  • Page 37 <6 mm Tighten the set screw using the 1.5 mm Hex wrench provided with the Amersham ECL Gel Box, to lock the expandable plug of the adapter into the socket. Make sure that the adapter is locked into the socket.

  • Page 38: Protocols

    5 Protocols Protocols About this chapter This chapter presents protocols for PAGE using Amersham ECL Gel Box and Amersham ECL Gel. Additionally, protocols for post-staining and transfer are provided. In this chapter This chapter contains the following sections: Section page 5.1 Electrophoresis protocol...

  • Page 39: Electrophoresis Protocol

    5 Protocols 5.1 Electrophoresis protocol Electrophoresis protocol In this section This section describes how to perform PAGE using Amersham ECL Gel Box and Amersham ECL Gel. Precautions WARNING Do not use Amersham ECL Gel Box if it is not working...
  • Page 40 Rinse the gel cassette with distilled water. Peel off the tapes from the two legs of the cassette. Place Amersham ECL Gel in Amersham ECL Gel Box so that the well side of the cassette faces toward the cathode (-) and the other cassette leg faces toward the anode (+).
  • Page 41 5 Protocols 5.1 Electrophoresis protocol Step Action Wiggle the comb back and forth, and bring it straight up from the cassette to make the wells available for sample loading. Note: Small gel pieces can be detached from the well container. This will not affect gel performance.
  • Page 42 Note: To ensure uniform mobility, load an equal volume of 1× SDS sample buffer into any unused well. Place the safety lid on top of Amersham ECL Gel Box. Running conditions Run the gel according to the recommendations below. Parameter...
  • Page 43 5 Protocols 5.1 Electrophoresis protocol Parameter Value Run time 60 minutes WARNING Do not exceed the maximum operating voltage of 200 V Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...

  • Page 44: Removing The Gel From The Cassette

    Once the run is completed, shut off the power, disconnect the electrodes, remove the safety lid and finally, remove the gel cassette from Amersham ECL Gel Box. Open the cassette by inserting the edge of the comb in the slot opposite the sample wells, and twist.
  • Page 45 5 Protocols 5.2 Removing the gel from the cassette Step Action Cut the stacking gel with the end of the top plate approximately 2 cm downstream of the wells. Repeat the procedure on the other side of the gel to remove the front. Hold the gel cassette bottom plate over a container with suitable buffer, with the gel facing downwards.

  • Page 46: Post-Staining Of The Gel

    Post-staining of the gel In this section All standard staining procedures, including Coomassie staining, Silver staining and Deep Purple™ staining, may be used with Amersham ECL Gel. For staining solutions recommended by GE Healthcare, see Gel staining reagents, on page Coomassie staining...
  • Page 47 5 Protocols 5.3 Post-staining of the gel Solution Preparation Preserving 1 Mix 25 ml of (87% v/v) glycerol with distilled water. solution 2 Make up to 250 ml with destaining solution. Coomassie staining protocol Step Action Fixation: Immediately after electrophoresis, immerse the gel in fixing solution for 30 minutes.
  • Page 48 5 Protocols 5.3 Post-staining of the gel Deep purple staining Introduction This section presents a Deep Purple staining protocol recommended by GE Healthcare. For more information refer to RPN6305 Deep Purple Total Protein Stain. Deep Purple staining solutions Solution Preparation Fixating/ 1 Mix 150 ml ethanol and 10 g citric acid (approx.
  • Page 49 Acidifying: Acidify gels for 45 minutes. Further washes in this solution can be performed if desired. Imaging: Imaging is best performed using laser scanning fluorescence flat bed imaging systems, such as the GE Typhoon™ FLA laser scanners. Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...
  • Page 50 (up to 6 months) add Deep Purple stain in 1:200 dilution. Silver staining Introduction This section presents a Silver staining protocol recommended by GE Healthcare. Refer to Silver Staining Kit Protein 71-7177-00 for more information. Silver staining solutions...
  • Page 51 5 Protocols 5.3 Post-staining of the gel Solutions Preparations Developing Mix 1 packet of sodium carbonate (6.25 g) and distilled • solution water. Make up to 250 ml with distilled water. • Note: Stir vigorously to dissolve the sodium carbonate Before use: Add 0.2 ml of formaldehyde (37% w/v).
  • Page 52 5 Protocols 5.3 Post-staining of the gel Step Action Washing: Remove the Staining solution. Rinse three times in distilled water for one minute each time. Developing: Add developing solution and gently shake for 3 minutes. Transfer the gel to stopping solution when the bands/spots have reached desired intensity.

  • Page 53: Transfer Protocol

    20 seconds, then in distilled water for 5 minutes and finally, in cold transfer buffer for at least 10 minutes. Note: Use a pre-cut membrane provided by GE Healthcare. See Chapter8 Related products, on page Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...
  • Page 54 5 Protocols 5.4 Transfer protocol Preparing the transfer tank Step Action Fill the transfer unit with transfer buffer. For information on how to use the transfer unit, refer to the instructions provided by the manufacturer of the transfer unit. Place the open transfer cassette with the anode side (+) down in a tray with transferbuffer and ensure that it is immersed in at least 3 cm of the transfer buffer.
  • Page 55 5 Protocols 5.4 Transfer protocol Step Action Layer 2: Place the two pre-wetted blotting papers on the sponge and press gently to remove any air bubbles. Note: Use a pipette or stirring rod to gently roll out any air bubbles trapped while assembling the stack.
  • Page 56 Please refer to 28-4025-91 for detailed instructions for use of Semi-dry blotters TE70 and TE77. Post transfer After transfer, continue with Western blotting detection or Post-staining detection. For information regarding GE Healthcare recommended products, Chapter 8 Related products, on page Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...

  • Page 57: Maintenance

    WARNING Cleaning. Do not autoclave, bake or microwave Amersham ECL Gel Box. WARNING Cleaning. Do not wash Amersham ECL Gel Box in a mechanical washer. WARNING Cleaning. Do not use abrasive creams or scourers. NOTICE Cleaning.
  • Page 58 Let the instrument air dry, or, wipe with a soft damp tissue. Keep power cables and connectors dry. Storage Store Amersham ECL Gel Box in a dry location at room temperature. Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC...

  • Page 59: Troubleshooting

    7 Troubleshooting Troubleshooting Problems Possible causes Remedies Low or no current Interrupted circuit Add some running buffer to • during the run each tank of Amersham ECL Gel Box Ensure that the tape is • removed from the gel cassette Ensure that the lid is in place •...
  • Page 60 7 Troubleshooting Problems Possible causes Remedies Bands difficult to Incorrect gel selection, sample Select a gel that separates in • distinguish overloading the desired molecular weight range Reduce loaded protein • amount For proteins of similar • molecular weight, a 2-D separation may be required Sample spreading Excess salt in the sample...
  • Page 61 7 Troubleshooting Problems Possible causes Remedies No green light, low Incomplete circuit Check all power connections • current (<20 mA) Check buffer level in tanks • at start. Ensure that the gel cassette • is in place Ensure that there is no tape •...

  • Page 62: Related Products

    8 Related products Related products In this chapter This chapter presents a subset of related products. For more information, refer to www.gelifesciences.com. Sample preparation Product Quantity Code no. SDS-PAGE Clean-up kit 50 samples 80-6484-70 Mammalian Protein Extraction Buffer 1 (for 500 ml) 28-9412-79 2-D Quant Kit 500 assays...
  • Page 63 8 Related products Power supplies Product Quantity Code no. EPS 301 Power Supply 18-1130-01 Amersham ECL Gel Product Quantity Code no. Amersham ECL Gel 10%, 10 wells 28-9898-04 Amersham ECL Gel 12%, 10 wells 28-9898-05 Amersham ECL Gel 4-12%, 10 wells 28-9898-06 Amersham ECL Gel 8-16%, 10 wells 28-9898-07...
  • Page 64 8 Related products Product Quantity Code no. Amersham ECL Gel 10%, 10 wells 28-9898-08 Amersham ECL Gel 12%, 10 wells 28-9898-09 Amersham ECL Gel 4-12%, 10 wells 28-9901-51 Amersham ECL Gel 8-16%, 10 wells 28-9901-52 Amersham ECL Gel 4-20%, 10 wells 28-9901-53 Electrophoresis buffers Product...
  • Page 65 8 Related products Transfer equipment Product Quantity Code no. TE 70 PWR Semi-Dry Transfer Unit, 14 x 16 cm 11-0013-41 TE 77 PWR Semi-Dry Transfer Unit, 21 x 26 cm 11-0013-42 Amersham ECL Multiprobe 11-0033-95 Amersham ECL Multiprobe XL 11-0033-96 TE 22 Mini Tank Transfer Unit 80-6204-26 TE 62 Transfer Unit...
  • Page 66 8 Related products Blotting membranes Product Quantity Code no. Blotting paper 3 MM Chr 20 x 20 cm 100 sheets 3030-861 Blotting sandwich Protran™ BA83, 0.2 μm blotting sandwich 10 units 28-9909-83 (8 × 7.5 cm) Protran BA85, 0.45 μm blotting sandwich 10 units 28-9909-93 (8 ×...
  • Page 67 8 Related products Amersham ECL HRP-linked Secondary Antibodies Product Quantity Code no. ECL Mouse IgG, HRP-Linked Whole Ab (from sheep) 1 ml NA931-1ML ECL Human IgG, HRP-Linked Whole Ab (from sheep) 1 ml NA933-1ML ECL Rabbit IgG, HRP-Linked Whole Ab (from donkey) 1 ml NA934-1ML ECL Mouse IgG, HRP-Linked F(ab)
  • Page 68 8 Related products Product Quantity Code no. Amersham ECL Western blotting reagents for 1000 cm RPN2109 membrane Amersham ECL Western blotting reagents for 4000 cm RPN2106 membrane Amersham ECL Western blotting reagents for 6000 cm RPN2134 membrane Amersham ECL Plex CyDye conjugated Antibodies Product...
  • Page 69 8 Related products Imaging Systems Product Quantity Code no. Typhoon Variable Mode Imagers Typhoon FLA 9500 29-0040-80 Typhoon FLA 7000 28-9558-09 ImageQuant Imagers ImageQuant™ LAS 4000 28-9558-10 ImageQuant LAS 4010 28-9558-11 ImageQuant LAS 4000 mini 28-9558-13 ImageQuant LAS 500 29-0050-63 Software and Accessories Product...
  • Page 72 All goods and services are sold subject to the terms and conditions of 751 84 Uppsala sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your Sweden local GE Healthcare representative for the most current information.

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