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2017

Labnet International,

Version # 2

ENDURO™

VE10 Vertical Gel Electrophoresis Systems

User Manual

ENDURO VE10 System

E2010-PA

ENDURO VE10 Electroblotting System

E2010-PBA

Lit #M00608

Содержание ENDURO E2010-PA

Страница 1: ...Copyright 2017 Labnet International Version 2 ENDURO VE10 Vertical Gel Electrophoresis Systems User Manual ENDURO VE10 System E2010 PA ENDURO VE10 Electroblotting System E2010 PBA Lit M00608...

Страница 2: ...cking Lists 3 Care and Maintenance 5 Setting Up 6 Gel Casting 8 Gel Preparation 8 Gel Selection 9 Gel Pouring 10 Sample Preparation and Loading 11 Gel Running 12 Blotting Insert Setup 14 Blot Running...

Страница 3: ...y interlock to the user When the lid is removed the current to the unit is broken DO NOT attempt to use the unit without the safety lid correctly positioned Always turn the power supply off prior to r...

Страница 4: ...tanks trays and other parts The tanks should be thoroughly rinsed with warm or distilled water but vigorous cleaning is not necessary or advised Air drying is recommended before use Cleaning Products...

Страница 5: ...thick 12 sample combs E2110 PC E2110 CP E1107 EP ENDURO VE10 Electroblotting System Units include tank lid internal module electrodes Blotting module and include the following accessories Glass Plate...

Страница 6: ...UTION DEGREE 2 in accordance with IEC 664 POLLUTION DEGREE 2 states that Normally only non conductive pollution occurs Occasionally however a temporary conductivity caused by condensation must be expe...

Страница 7: ...Always wear gloves and safety glasses RNaseZAP Ambion can also be used Please consult the instructions for use with acrylic gel tanks Questions and Service Should you have a question about the operat...

Страница 8: ...nd then with 70 ethanol One set of glass plates constitutes one notched glass plate and one plain glass plate with bonded spacers When using a triple glass plate sandwich two notched glass plates one...

Страница 9: ...e sandwich the pressure bars will need to be in the completely open position 4 Position the ENDURO VE10 Gel module on a flat surface Do not insert the ENDURO VE10 Gel module into the casting base at t...

Страница 10: ...7 With the cam handles facing directly downwards rotate the cams in the opposite direction 1800 or until the insert has tightened down onto the gasket Do not overturn as this will cause the glass plat...

Страница 11: ...s of other thickness are to be run adjustments will need to be made 1 Put together bonded spacer plain glass plate with notched plate 3A Fully tighten screws ensuring not to wobble unit 2 Insert insid...

Страница 12: ...ty given different sizes of protein See Table 2 below which details which percentage of gel to use to separate the sizes of proteins indicated Table 2 Acrylamide Percentage Separating Resolution 5 60...

Страница 13: ...s is the level for the resolving gel 2 Add 15 l of TEMED to the resolving gel solution 3 Fill the glass plates to the line and avoid generating any air bubbles Filling must be performed quickly before...

Страница 14: ...inhibit sample progression 11 Allow the stacking gel polymerize for 30 minutes For continuous gels 4 Follow the instructions for mixing the acrylamide solution Add 15 of TEMED and mix well but avoid...

Страница 15: ...or 20 seconds at 12 000 rpm The protein samples are now ready to load Loading the samples 1 If desired fit the previously frozen cooling pack s into the tank The longest side of the ice pack should be...

Страница 16: ...a pipette tip taking care not to damage the wells or induce any air bubbles 6 Fill any unused wells with 1X sample buffer 7 It is a good idea to note the orientation and order the samples were loaded...

Страница 17: ...225V 40 90mA Three gels 90 225V 60 135mA Four gels 90 225V 80 180mA The recommended power condition for optimal resolution with minimal thermal band distortion is 150 volts constant voltage setting N...

Страница 18: ...or other suitable surface b Pre soaked fiber pad c Two pieces of 45 m filter paper pre soaked in buffer d Gel cut the left hand corner for indexing e Transfer membrane Follow manufacturer s direction...

Страница 19: ...o the slots in the module with the black side of each adjacent to the negative electrode It is a good idea to note the orientation and order in which the blot sandwiches were loaded 2 Use of a magneti...

Страница 20: ...of each cassette and gently pry apart the blot sandwich and remove the membrane from the gel 8 The membrane can now be further processed Remember to save the filter paper behind the gel to check for...

Страница 21: ...Water Stock 4X Stacking Tris 0 5 M Tris HCL pH 6 8 0 4 SDS To 110 mL Distilled Water add 12 12 g of Tris base Add 8 mL of 10 SDS Adjust pH to 6 8 with 1N HCl Adjust the final volume to 200 mL with Di...

Страница 22: ...is Nylon Microporous membrane modified with strongly basic charged groups Binds negatively charged macromolecules DNA or RNA with low background Pore Size 45 m Can Re probe Southern Transfer Northern...

Страница 23: ...els Towbin Buffer pH 8 3 25 mM TRIS 192 mM glycine 20 Methanol 3 0 gm TRIS 14 4 gm glycine 200 ml Methanol add ddi H2O to 1 liter Denatured Gels Towbin Buffer pH 8 3 no Methanol 25 mM TRIS 192 mM glyc...

Страница 24: ...A 1 800 492 1110 Canada 1 978 442 2200 For other regions of the world please visit www corning com lifesciences or see the included instruction manual for a list of World Wide Support Offices Corning...

Страница 25: ...0 x 10 cm E2110 NG 1 BS Notched Glass Plate with bonded 1 mm spacers 10 x 10 cm E2110 NG 1 5 BS Notched Glass Plate with bonded 1 5 mm spacers 10 x 10 cm E2110 NG 2 BS Notched Glass Plate with bonded...

Страница 26: ...labnetinfo corning com www labnetinternational com LN135000...

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