Membrane Protein Crystallization Plates

From: Yuri Pompeu
Date: 25 February 2012 20:35


Hello Everyone,
I am considering the purchase of crystallization plates for membrane proteins.
I would love to hear what some of the community thinks or has experienced with these.
Particulalrly the monoolein and monoolein/cholesterol coated plates ( I am not sure I can mention the vendor here but it "should" not matter)
So fire away. Is it worth it? Any succes stories? Bad experiences?
I appreciate the input
Best,
Yuri

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From: Pius Padayatti


Hi Yuri,
i strongly suggest these plates sold through Hampton research
Paul Marienfeld GmbH plates for your use.
here is a link , best plates in the market.
link: http://hamptonresearch.com/product_detail.aspx?cid=10&sid=182&pid=611

Plates comes with extra cover slips in addition to single glass cover
for four wells at a time
whcih let you set these plates manually if you like it that way.

Good for automations as well.
only problem: since the galss is siliconized you can not write anything on it
(may be there is some industry grade marker might work.)
But there is enough place on side to stick a printed label or bar code
for your'automation
for keeping records
hope this is what you wanted to know.

if you are money crunched and want to make some plates
on your own. buy regular micoscope slides, some two sided
tapes (3M) very thin and a tool that sold for cutting out small pieces
of tissue samples. it looks like a pen with sharp
punch enough to make a hole in a 3M tape
make a 27 hole plate in a standard microslide.

cheers
pius

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From: Pius Padayatti


Hi board,
Although i do not want to endorse anything personally
earlier mail to Yuris response reflects my personal reflection of what i like.

Here is a plate from Molecualr dimensions that i found
may be an alternate where you can chose the sandwich thickness
good idea but do not know what advantage it might have on
crystallization process.

link:
http://www.moleculardimensions.com//shopdisplayproducts.asp?Search=Yes

Thanks to Tony from molecular dimensions who point this out.
quote:
"We have many users of Laminex which offers a lot of flexibility
in that you can choose sandwich thickness as well as glass or plastic bases
and glass, plastic or film covers.
If you have not tested Laminex I would be pleased to send you a sample trial
pack.

Tony Savill
Molecular Dimensions Inc."

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From: Patrick Shaw Stewart

Yuri

I know this isn't quite what you're asking, but it's helpful to use the COC ("UV permeable") version of plates if you're crystallizing samples that contain detergent.  It's just that the drops tend to spread very thinly if you use the normal polystyrene PS plates.  The COC is more hydrophobic.  (On the other hand we prefer PS plates for normal proteins with no detergent.)

Patrick

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From: Pius Padayatti


Hi
 i do not want to get into trouble by going against any products
The plates what you are talking about as soon it came out we tested.

I did not look back to see if there was a monoolein:cholestrol coated plate.
The ones i used were for sure monoolein coated ones.
That tells it all.The rest of the plates still unused.

I set up plates with bacteriorhodopsin even that did not work in our hands
(the bacteriorhodopsin set up otherwise would
work very well.)

By the way it did not work for me did not mean it is bad.

And the plates were in plasticware made it highly birifringent to
document the results.

But the plates Patrick says here i am not sure about.
If Patrick want to to tell me a little bit more about i would like to hear that.

I would set up plates the normal way.
I am really not convinced mesophase will form
by just adding protein-detergent to a previously poured monoolein.
anyway i am also interested to hear experts views of the method
adapted in this product.
There is a poster that they shown at their website shows
bacteriorhodopsin and another membrane protein crystallized using these
plates.

These are my personal opinion.

Pius

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From: Jim Fairman

Yuri,

Did you mean plates for setting up Lipidic Mesophases?  If so, here is a listing of products I have used in the past.  I highly endorse the plates from Molecular dimensions, particularly the plastic laminex plates (MD11-51-100 + MD11-54).  They do have the dis-advantage of drying out after a 2-month or so period, but they make the job of harvesting significantly easier than dealing with the glass equivalents (so much so that I don't ever plan on using the glass bases and covers again).

Molecular Dimensions:  Using the MD11-51-100 plastic bases combined with the MD11-54 covers results in a cost of $9.20 per plate.

Laminex Plastic 100 um base (MD11-51-100) - I've used these to solve multiple structures during my post-doc at NIH.  They're UV transparent for doing UV microscopy and detection of your crystals.  Cost: $66 USD for a pack of 10.

Laminex Plastic 200 um base (MD11-50) - Has a thicker 200 um layer for inputting larger amounts of mesophase and precipitant.  Cost:  $66 USD for a pack of 10.

Laminex Film Cover (MD11-54) - Used to cover the top of the "sandwich" plate.  Also UV transparent.  Cost: $26 USD per pack of 10.

Laminex Glass 100 um base (MD11-50-100) - Glass base with 100 um spacer. Cost:  $66 USD per pack of 10.

Laminex Glass 200 um base (MD11-50) - Glass base with 200 um spacer.  Cost: $66 USD per pack of 10.

Glass Covers (MD11-52) - Glass for forming sandwich plates.  Cost: $40 per pack of 10.

Hampton Research:

Plastic UV Transparent LCP Setup Kit (HR3-186) - Set of 20 plastic plates in a slightly different form factor than the Mol Dim. plastic plates.  Cost: $483 USD per pack of 20.  This is $24.15 per plate.

Glass UV Transparent LCP Setup Kit (HR3-151) - Siliconized glass bases and glass tops.  Cost: $368 USD per pack of 20 ($18.40 per plate). 

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From: Pius Padayatti


Quote"Particulalrly the monoolein and monoolein/cholesterol coated plates

( I am not sure I can mention the vendor here but it "should" not matter)"

Since the person who asked this question here
forget about it alltogether to write something back
here is what he was asking about (i think)

Anybody heard about the nextal plates
see the link
http://www.qiagen.com/products/nextalcubicphaseproducts.aspx

It uses a different method of mixing lipids other than
using the syringe method.
They have lipid layered on the bottom of the well
and protein samples are pipetted
onto the monoolein.and later on precipitant.

Plastic based ones are really bad idea
Not everyone have an access to UV
even if UV screening is used not a full-proof method
to detect membrane protein micro crystals

I would definitely think glass based plates are the way to go.

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From: Yuri Pompeu

Thanks for all the replies.
I will try a couple of different plates/set-ups. My favorite will be the one that gives me a crystal ;)