Reason and Application
Expedient exchange for high sub-atomic weight proteins (200 kDa <)
MeOH, Acid, Detergent free
Highlights
Powerful exchange of high particles (up to MW500 kDa)
An opportunity to finish smearing is 30-60min, Greater time shortening
5x concentrated reagent-Dilute with refined water before use
MeOH, Acid, Detergent free
For 1 year at room temperature in dim
An example is accessible. If it’s not too much trouble, demand the example through the “Reach Us” menu at the top.
Information
While this smearing arrangement is utilized, it required 250-450mA per 1 Mini gel sheet. So we suggest a power supply of high current result type for it. If it’s not too much trouble, reach us for additional subtleties.
1. Wellbeing safeguards
Prior to utilizing the item, read this manual
completely from the outset. Try not to begin the activity until
you get the items in manual. This
report makes sense of just strategies used for
indicated purposes. Try not to involve the item for any
reason or by some strategy not portrayed in the
manual. In the event that it is utilized for any reason or by any
technique not depicted in the manual, an administrator
ought to take more time for all expected security
measures and possibilities. Likewise, read a manual of
hardware utilized with it all the while.
2. Application reason
The item is support to move proteins from a gel
to a film by semi-blotching strategy later
electrophoresis.
5. Safeguarding technique
● Keep unopened EzFastBlotHMW at room
temperature.
● It is steady until the termination date for use
(indicated on the mark) in the event that it isn’t opened.
● The functioning arrangement is likewise storable at room
temperature. However, go through it quickly.
6. Removal strategy
● Follow a removal technique laid out by the association you have a place with.
● Materials of jug Fundamental body:High thickness polyethylene
Top :Polypropylene
7. Important things other than the iteme.g., the accompanying items are expected for
smearing of a polyacrylamide gel of Mini size
(85x90x1mm thick).
● A layer
(WSE-4051 P in addition to layers
PVDF 85mm x 90mm )
● 6 channel papers
(CB-09A Filter papers 85mm x 90mm x
0.8mm)
● A semi dry smudging framework
(WSE-4020 HorizeBLOT 2M-R)
(WSE-4040 HorizeBLOT 4M-R)
● A power supply
(WSE-3200 powerStation ⅲ )
(WSE-3500 powerStation HC )
8. Safety measures for use
● The item is 5x stock arrangement. Weaken it to the
thickness of 1/5 with refined water, when it is
utilized.
● The smudging gadgets of our old model cutoff max
electric flow (Ampere) as follows.
AE-6687 HorizeBLOT 2M :1.0a
AE-6677 HorizeBLOT:1.0A
AE-6688 HorizeBLOT 4M :1.2A
AE-6678 HorizeBLOT :1.2A
On the off chance that you utilize higher current than the abovementioned, the
gadget might be broken.
HorizeBLOT 4M whose parcel number is later
490011 can give electric flow till 2.4A.
Likewise, a gadget of other organization might restrict max
current and voltage.
Affirm the determination before an activity.
9. Utilization
A.Dilute reagent
To make the functioning arrangement (blotching support), weaken
the item to the thickness of 1/5 with refined water.
B.Make a smearing film hydrophilic
Assuming a PVDF film is utilized for smudging, make it hydrophilic by the accompanying technique ahead of time.
1. Empty a few mL methanol into a holder
greater than a PVDF film. Submerge it in
the methanol for around 10 seconds.
2. Pour around 50 mL smudging support into another
compartment. Take the layer out from
methanol and placed it in the smearing cushion. Shake
it for 5~15 minutes. It will in general drift at the
starting. Shake it to drench in the arrangement
totally.
C.Blotting
1. Get ready 6 channel papers (CB-09A). Assuming you use channel
papers of other organization, slice it to a similar size
as a gel, and set up the quantity of it to be
in excess of 6 mm thick altogether.
Pour around 50 mL smudging support into a
holder greater than it, and submerge it in the
arrangement. (Refer to conditions on the below.)
2. After electrophoresis, put a gel in around 50 mL
smearing cradle, and wash the surface daintily
(in no less than a few minutes) to dispose of little offers
of gel and air pockets of SDS. Try not to drench or
shake it for quite a while.
3. Heap channel papers, the layer and the gel with
alluding to the above figure.
① Drop a few mL blotching cradle into the lower
anode plate to get wet ahead of time.
② Put 3 channel papers inundated in smearing cushion
on the cathode plate.
③ Put the layer on the channel papers.
④ Drop a few mL smudging cushion into the
layer.
⑤ Put the gel on the layer without embedding
air rises between them.
⑥ Put 3 channel papers drenched in blotching support
on the gel.
⑦ To improve attachment of the gel and film,
push out superfluous support with a roller
connected to the gadget. Or on the other hand, push a few spots of
the heap emphatically and equitably from above by
hands wearing gloves. On the off chance that the attachment is
lacking, grinning or lopsided smudging productivity
may happen.
⑧ Set the upper anode plate on the channel
papers smoothly. Associate the gadget and a power
supply with lead wires. The lower anode of
HorizeBLOT 2M-R/2M is anode (+). Associate the
red terminal of lead wires to the red connector
of force supply.
⑨ Apply 25V of steady voltage for 15~30
minutes on account of WSE-4020/4040.
Hand-made gel (Compact/Mini size gel): 25V
Precast gel (Compact/Mini size gel): 25V
Set current worth as around 250~450 mA per a
gel around then.
Apply 6~7 mA of consistent current per 1 cm2
for 15~30 minutes on account of AE6687/6688.
Minimized gel(60mm x 60mm):250mA/gel
Small size gel(85mm x 90mm):450mA/gel
Set voltage esteem as 25~40V around then. The
voltage ought to be set as 40V or less.
In the event that current can not be set as the abovementioned, change
an opportunity to apply it. For instance, if 225 mA of
electric flow streams in a Mini size gel, the time is
30~60 minutes.
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If WSE-4110 powered BLOT-ONE which is implicit
power supply unit is utilized with the item, read the
guidance manual of the gear completely.
Assuming the item is utilized with AE-6685 poweredBLOT
scaled down, it tends to be utilized under a similar state of
EzBlot.