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22 

 

Appendix 

A-1. Buffer recipes 

eZwest cleaning solution (0.1M NaOH, 10% ethanol) 

Recipe 
NaOH

 

4g 

Ethanol

 

100 ml 

Add water to

 

1L 

 

Mix thoroughly 

 
Regular wash buffer 

Recipe

 

1

×

 PBST / TBST

 

1L 

 
10x TBS buffer 

Recipe

 

Tris base 

 

24g 

NaCl  

88g 

 

Dissolve in 900mL water, pH to 7.4 
with 12N HCl 

Add water to 

1L 

 
1x TBST buffer 

Recipe

 

10

×

 TBS

 

100mL 

Tween20

 

0.5mL 

Add water 

900mL 

 

Mix thoroughly, pH to 7.4 

 
10x PBS buffer 

Recipe 
Na2HPO4·2H2O 

17.8 g 

NaCl 

80g 

KCl 

2g 

KH2PO4 

2.4g 

Add water to 

1 L 

 

Mix thoroughly 

 

Содержание eZwest Lite

Страница 1: ...Automated Western Device...

Страница 2: ......

Страница 3: ...ons 8 2 1 Instrument display and feature location 8 2 2 Basic operation 10 Save cassette ID when using the cassette for the first time 10 View and adjust method 11 Set cassette specific method 14 2 3...

Страница 4: ...2...

Страница 5: ...set up the device before your first experiment Note Before relocating the instrument please drain the system completely Step Description 1 Check and make sure the power plug matches the outlet 2 Plac...

Страница 6: ...tion done by any other party than GenScript or an authorized agent Use of fittings or other spare parts supplied by any other party than GenScript Damages caused by disasters For consultation and main...

Страница 7: ...iability typically associated with manual operations contributing to experimental reproducibility The eZwest Lite device is compatible with the traditional reagents and protocols used for conventional...

Страница 8: ...e size WxH 90x83 mm 3 5x3 2 in Throughput 1 channel for up to 2 membranes Application Western blotting eZwest Lite ABS PP stainless steel silicone Forceps Stainless Steel Roller PC 1 3 Ordering inform...

Страница 9: ...west device extensive cleaning After every 20 use or every 2 weeks perform the extensive cleaning following the procedures below 1 Empty the Wash bottle and Waste bottle 2 Add 100 mL deionized water i...

Страница 10: ...8 2 Instructions 2 1 Instrument display and feature location eZwest Lite automated western device Fig 1 Front view Fig 2 Back view...

Страница 11: ...9 Fig 3 Top view Fig 4 Membrane cassette Fig 5 Display main menu...

Страница 12: ...shown in Fig 7 2 Switch on the device insert the membrane cassette into the device Make sure the pins are pushed all the way in and the side with the eZwest logo is upright Note The pins are designed...

Страница 13: ...ress OK to save cassette ID as show in Fig 8 Note Each eZwest Lite device could save up to 5 cassette IDs Fig 8 View and adjust method 1 Enter method setting by pressing Method on the upper right corn...

Страница 14: ...to show and modify the associated details Note Each Method may be programmed with 7 steps Fig 11 4 Each step within a Method is customizable a Reagent option Fig 12 b Number of Cycles required to repe...

Страница 15: ...13 c Run time for each cycle in minutes Fig 14 d Pump Speed for recirculating the liquid flow through the membrane cassette Fig 15 5 Once you have finished setting up your Method press Save...

Страница 16: ...Change name of membrane cassette a Under parameter interface select the name of membrane cassette Fig 16 b Press Edit ID and change name with the pop up keyboard Fig 17 2 3 Using the Pre programmed b...

Страница 17: ...e the latch by rotating it up as shown in Fig 19 i lift the latch and pull it towards the top of the cassette as in Fig 19 ii the opened cassette is shown in Fig 19 iii Fig 19 i ii iii 4 Placing membr...

Страница 18: ...of the first membrane along the membrane edge Lay the second membrane on top with the protein side facing down and high MW side align towards the bottom edge of the cassette Make sure both spacers are...

Страница 19: ...result in experiment failure 8 Select the method on the main menu and click START to run 9 Once method finish click FINISH and replace the membrane cassette with cleaning cassette Membranes are ready...

Страница 20: ...min Normal Wash 4 3 min Normal Normal Protocol All steps performed at room temperature Step Cycles Time Pump Speed Blocking 1 60 min Normal Wash 5 3 min Normal 1 Antibody 1 120 min Normal Wash 5 3 min...

Страница 21: ...with primary antibody overnight at 4 C with shaking The following day insert membrane into eZwest and run the following method at room temperature Step Cycles Time Pump Speed Wash 5 3 min Normal 2 An...

Страница 22: ...trong background Insufficient wash 1 Increase number of wash cycles set to 5 cycles for 3 minutes each 2 Increase pump flow per wash cycles set to normal Weak signal 1 Antibody is too diluted 2 Instru...

Страница 23: ...lations citations handbooks etc 2 Complete technical support contact information 3 Access to the GenScript Online Catalog 4 Additional product information and special offers For more information or te...

Страница 24: ...gular wash buffer Recipe 1 PBST TBST 1L 10x TBS buffer Recipe Tris base 24g NaCl 88g Dissolve in 900mL water pH to 7 4 with 12N HCl Add water to 1L 1x TBST buffer Recipe 10 TBS 100mL Tween20 0 5mL Add...

Страница 25: ...23 1x PBST buffer Recipe 10 PBS 100mL Tween20 0 5mL Add water 900mL Mix thoroughly pH should be around 7 4...

Страница 26: ...24...

Страница 27: ......

Страница 28: ...GenScript USA Inc 860 Centennial Ave Piscataway NJ 08854 Tel 732 885 9188 1 877 436 7274 FAX 1 732 5185 5150 Email product genscript com...

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