Hoefer SE615 User Manual
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Hoefer SE615 User Manual

Multiple gel casters
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Hoefer SE615
and SE675
Multiple gel casters
 
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SE615-IM

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Summary of Contents for Hoefer SE615

  • Page 1 Hoefer SE615 and SE675 Multiple gel casters   SE615-IM...

  • Page 2: Table Of Contents

    Page finder Safety warnings and precautions ......viii Introduction ............1 Setting up the caster ..........5 Casting with divider plates ........7 Pouring standard homogeneous gels ......9 Pouring standard gradient gels ......11 Pouring stacking gels ...........15 Removing polymerized gels ........17 Troubleshooting ...........18 Care and maintenance .........20 Appendix A: Gel identification numbers ....21 Appendix B: References ........21 Ordering information ..........22...

  • Page 3: Safety Warnings And Precautions

    Safety warnings and precautions • Acrylamide is a neurotoxin. • Wear gloves when handling acrylamide or polyacrylamide. • Wear a dust mask when weighing acrylamide or preparing acrylamide solutions. Protecting your equipment To keep your instrument in excellent condition, please take the following important steps: •...

  • Page 4: Introduction

    SE600 Chroma Vertical Slab Gel Electro- phoresis Units. Homogeneous and gradient gels The SE615 and SE675 can be used for casting both homogeneous and gradient gels. Gradient gels are cast through the port at the bottom of the caster. When casting homogeneous gels, the results are most consistent when the gels are cast from the bottom.
  • Page 5 The maximum number of gels you can cast simultaneously using the standard casting proce- dure is shown in Table 1. Table 1. Maximum number of standard gels cast 0.75-mm  1.0-mm  1.5-mm  spacers   spacers  spacers SE615 SE675 •...
  • Page 6 0.75-mm  1.0-mm  1.5-mm  spacers   spacers  spacers SE615 SE675 See Fig 1 on page 4 to identify the components of the multiple gel casters. For information on parts, accessories and related equipment, see page 22. Pouring second-dimension slab gels...
  • Page 7 Fig 1. Components of the SE615 and SE675 Multiple Gel Casters. SE615 and SE675 components no.  description Caster body, includes faceplate Spring clamp (4) Foam cord gasket Silicone displacement block* Acrylic spacer block Comb Polycarbonate filler sheet Glass plate Spacer Inlet port Nylon thumb screw * A small silicone displacement block is also available for the SE675.

  • Page 8: Setting Up The Caster

    Setting up the caster Fig 2. Building a glass sandwich for caster standard casting. glass plate spacer one sandwich glass plate wax paper or polycarbonate sheet Preparing the caster If the faceplate is attached to the caster, remove it. Remove the four side clamps, loosen the two thumb screws, and slide off the faceplate.
  • Page 9 Building the stack Note:  To make separation of polymerized gel sandwiches Place a polycarbonate sheet in the caster so that 1/3 easier, place wax paper sheets of the sheet extends out of the top. in the caster between each set You can use this sheet as a lever when inserting filler of glass sandwiches.

  • Page 10: Casting With Divider Plates

    Casting with divider plates To double the number of gels run simultaneously on the SE600 and related units, place a notched glass divider plate and another pair of spacers between the standard glass plates. The result is a “club sandwich,” with two layers of gel formed by the three glass plates.
  • Page 11 Place a polycarbonate sheet or wax paper on top of the club sandwich. Repeat step 3, alternating wax paper and club sandwiches. Note:  For optimum visibility, After the last club sandwich, add acrylic spacer blocks place a polycarbonate sheet and extra polycarbonate sheets until the entire stack is against the faceplate.

  • Page 12: Pouring Standard Homogeneous Gels

    Table 3. Gel casters and volume required mask when weighing acrylamide or preparing acrylamide stocks. gel caster  required volume  gradient maker  (ml)  size* SE615 190–500 SG500 SE675 60 –150 SG100 or SG500 *Actual size depends on calculated volume. Prepare and degas enough monomer solution to fill all sandwiches to the level of the notched plate.
  • Page 13 Add initiator and catalyst, and pour the solution into Caution!  Isobutanol crazes the glass sandwiches from the top. or clouds the acrylic caster The solution flows from one sandwich to another walls, making it difficult to see through the groove at the bottom of the caster and the gels.

  • Page 14: Pouring Standard Gradient Gels

    Setting up the gradient maker Prepare a 50% glycerol displacement solution containing a small amount of bromophenol blue: approximately 75 ml for the SE615 and 35 ml for the SE675. Calculate the amount of gradient solution you will need by referring to the formula on page 9.
  • Page 15 Assemble the gradient components. (See Fig 5.) a. Close all gradient maker valves and place a stir bar in the mixing chamber, the one with the tube connector port. b. Attach one end of a piece of tubing to the outlet of the gradient maker.
  • Page 16 Pouring gradient separation gels Warning!  Acrylamide is a neurotoxin. Wear gloves Prepare and degas the two monomer solutions for the when handling acrylamide or gradient maker. polyacrylamide. Wear a dust Add glycerol or sucrose to the solution of higher mask when weighing acrylamide acrylamide concentration to stabilize the gradient as or preparing acrylamide stocks.
  • Page 17 When almost all the acrylamide solution is drained from the gradient maker, stop the pump and close the mixing valve. Tilt the gradient maker towards the outlet side and remove the last few milliliters of mix. Do not allow any air bubbles to enter the tubing. Add the glycerol displacement solution to the mixing chamber and start the pump.

  • Page 18: Pouring Stacking Gels

    Pouring stacking gels You may add stacking gels to the entire set of polymerized gels —homogeneous or gradient— while the sandwiches are still in the casting box, or you may add stacking gel to individual gels immediately prior to use. For optimal resolution, add the stacking gel at the last minute to prevent diffusion of buffers between the two gel layers.
  • Page 19 Replace the caster in a vertical position. Make sure Note:  Oxygen inhibits gel the surface of the separating gel is free of liquid. polymerization. Do not trap Use a pipette to fill each sandwich individually with air bubbles underneath the stacking-gel monomer solution.

  • Page 20: Removing Polymerized Gels

    Removing polymerized gels After the gels have polymerized, place the caster in a horizontal position. If you poured stacking gels, leave the combs in place. Remove the faceplate. Tip the caster to pour off the glycerol solution. Note:  The SDS buffer makes Slide the stack of sandwiches, acrylic blocks, and the glass plates slippery.

  • Page 21: Troubleshooting

    Troubleshooting symptom  possible cause  recommended action   1 . Gels adhere to glass plates  Dirt, grease or Soak plates in a strong laboratory detergent; when opening sandwich. fingerprints on plates. rinse well in distilled water. Handle with gloves only. 2. Gels cast simultaneously  Different amounts of Use the same amount of overlay on all are different sizes.
  • Page 22 symptom  possible cause  recommended action 8. No polymerization of  Insufficient APS Increase both APS and TEMED by 30–50% SDS gel (or incomplete  or TEMED. APS solution is old. Make up fresh APS polymerization.) each day. APS stack is wet. APS is hygroscopic. Open a fresh bottle. in gel solution. Degas at least 10 minutes.

  • Page 23: Care And Maintenance

    Care and maintenance • Do not autoclave or heat any part above 45 °C. • Do not expose the caster or its parts to organic solvents. Cleaning Rinse the caster, faceplate, silicone plugs and polycarbonate sheets in dilute detergent and rinse with distilled water.

  • Page 24: Appendix A: Gel Identification Numbers

    Appendix A: Gel identification numbers For positive identification of gels, label each slab by incorporating a small label printed on thin filter paper in the bottom corner of the gel. Use a carbon typewriter ribbon, photocopier or laser printer to make these labels, since many liquid- based inks are electrophoresed off paper during an SDS electrophoresis run.

  • Page 25: Ordering Information

    Ordering information product  quantity  code number SE615 Multiple Gel Caster Kit, 10 gels SE615 Includes 20 glass plates, space-saver plate, 5 filler sheets, 100 sheets of wax paper, and Spacer-Mate alignment template. (Order combs and spacers separately.) SE675 Multiple Gel Caster Kit, 4 gels...
  • Page 26 Combs  number  thickness   width      of wells  (mm)  (mm)  quantity  code number 0.75 SE511-10-.75 1.00 SE511-10-1.0 1.50 SE511-10-1.5 0.75 SE511-12-.75 1.00 SE511-12-1.0 1.50 SE511-12-1.5 0.75 SE511-15-.75 1.00 SE511-15-1.0 1.50 SE511-15-1.5 0.75 SE511-20-.75 1.00 SE511-20-1.0 1.50 SE511-20-1.5 0.75 SE511-28-.75 1.00 SE511-28-1.0 1.50 SE511-28-1.5 Comb depth 15 mm;...
  • Page 27 Hoefer, Inc. 32 Scotland Blvd, Ste. 9, Bridgewater, MA02324, USA Toll Free: 1-800-813-0488 Phone:1-508-807-4665 E-mail: support@hoeferinc.com Web: www.hoeferinc.com Hoefer is a registered trademark of Hoefer, Inc. © 2023 Hoefer, Inc. — All rights reserved.

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