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EVS1000 Blot units 

 

5

 

 

Protein (Western) blotting buffers 

Transfer buffers must be made accurately using high grade reagents. Do not adjust the pH 
with acid or base as this will affect the properties of the buffer. pH will vary according to the 
purity of the reagents used.  

Triple buffer system 

For high efficiency transfer of Protein from acrylamide gels  

 

Anode 1 Buffer:  
0.3 M Tris base, 20% MeOH, pH 10.4:  
soak 4 standard grade filter paper sheets 

 

Anode 2 Buffer:  
0.025 M Tris base, 20% MeOH pH 10.4:  
soak 2 standard grade filter paper sheets 

 

Cathode Buffer:  
0.025 M Tris base, 0.04 M Caproic Acid, 20% MeOH pH 9.4:  
soak 6 standard grade filter paper sheets 

DNA (Southern) Blotting Buffer 

For high efficiency transfer of DNA from agarose gels. 

 

Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in: 

o

 

50 x 1 M ethanolamine-glycine buffer, pH 11 

RNA (Northern) Blotting Buffer 

For high efficiency transfer of RNA from agarose gels. 

 

Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in: 

o

 

50 x 0.2 M morpholinopropanesulfonic acid (MOPS) 

o

 

50 mM sodium acetate 

o

 

5 mM EDTA pH 7.0 

Nucleic acid transfer buffers 

Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in: 

 

1 x TAE  40 mM Tris (pH 7.6), 20 mM acetic acid, 1 mM EDTA 

 

50 x (1l) dissolve in 750 ml distilled water: 

o

 

242 g Tris base (FW = 121) 

o

 

57.1 ml glacial acetic acid 

o

 

100 ml 0.5 M EDTA (pH 8.0) 

 

Fill to 1 litre with distilled water. 

Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in: 

 

1 x TBE 89 mM Tris (pH 7.6), 89 mM boric acid, 2 mM EDTA 

 

10 x (1l) dissolve in 750 ml distilled water: 

o

 

108 g Tris base (FW = 121) 

o

 

55 g boric acid (FW = 61.8) 

o

 

40 ml 0.5 M EDTA (pH 8.0) 

 

Fill to 1 litre with distilled water. 

Transfer buffers 

Towbin Buffer with 20% methanol 

Soak 12 pieces of standard grade filter paper, 6 for the cathode, 6 for the anode in: 

 

0.025 M Tris base 

 

0.192 M glycine 

 

20% methanol 

 

pH 8.3 

 

 

Содержание EV1300

Страница 1: ...EVS1000 BLOT series MANUAL Rev 44 2019 ...

Страница 2: ......

Страница 3: ...tenance 1 Environmental conditions 2 Fitting electrode cables 2 Electroblotting 3 Setting up the blot sandwich 3 Blot run conditions 3 Buffer solutions for blotting 4 Protein Western blotting buffers 5 Nucleic acid transfer buffers 5 Transfer buffers 5 Certificate 6 Warranty 7 ...

Страница 4: ...s of various gases are produced at the electrodes The type of gas produced depends on the composition of the buffer employed To disperse these gases make sure that the apparatus is run in a well ventilated area General care and maintenance Units are best cleaned using warm water and a mild detergent Water at temperatures above 60 C can cause damage to the unit and components The inner module shoul...

Страница 5: ...tion occurs Occasionally however a temporary conductivity caused by condensation must be expected Fitting electrode cables Note Before setting up the tank please ensure that it has been properly cleaned and dried Note the position of the lid on the unit This shows the correct polarity and the correct orientation of the cables black is negative and red positive Remove the lid from the unit Note If ...

Страница 6: ...assette 1380 ml 2800 ml 5600 ml Two cassettes 1290 ml 2620 ml 5240 ml Three cassettes 1200 ml 2440 ml 4880 ml Four cassettes 1110 ml 2260 ml 4520 ml Each cooling pack will take the place of 100 ml of buffer for EVS1100 and 500 ml of buffer for EVS1200 and EVS1300 Blot run conditions Insert the cassettes into the slots in the module with the black side of each adjacent to the negative electrode It ...

Страница 7: ...mbrane from the gel The membrane is now ready to be probed Duration of blotting EVS1100 EVS1200 EV1300 One hour 100 V 400 mA 100 V 400 mA 100 V 400 mA Three hours 50 V 200 mA 50 V 200 mA 50 V 200 mA Buffer solutions for blotting Do not adjust the pH when making these buffers as this will cause blot over heating The pH will vary according to the freshness of the reagents used Towbin Buffer 25 mM Tr...

Страница 8: ...orthern Blotting Buffer For high efficiency transfer of RNA from agarose gels Soak 12 pieces of standard grade filter paper 6 for the cathode 6 for the anode in o 50 x 0 2 M morpholinopropanesulfonic acid MOPS o 50 mM sodium acetate o 5 mM EDTA pH 7 0 Nucleic acid transfer buffers Soak 12 pieces of standard grade filter paper 6 for the cathode 6 for the anode in 1 x TAE 40 mM Tris pH 7 6 20 mM ace...

Страница 9: ...EVS1000 Blot units 6 Certificate ...

Страница 10: ...ce free of charge units where improper solutions or chemicals have been used For a list of these please see the Care and Maintenance subsection Consort products are for research use only Consort is not liable for consequential damages arising out of the use or handling of its products A return authorisation must be obtained from Consort before returning any product for warranty repair on a freight...

Страница 11: ...EVS1000 Blot units 8 ...

Страница 12: ...EVS1000 Blot units 9 Consort bvba Hertenstraat 56 unit 9 2300 Turnhout Belgium Tel 32 0 14 41 12 79 Fax 32 0 14 42 91 79 Sales sales consort be Support support consort be Information info consort be ...

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