[Histonet] RE: lab math: ihc dilution

[Truscott, Tom]

You are so right-that's why I always try to double check everything even my spellink. Tom T

-----Original Message-----
From: Elizabeth Chlipala [mailto:liz <@t> premierlab.com] 
Sent: Friday, November 21, 2014 12:05 PM
To: Truscott, Tom; 'histonet <@t> lists.utsouthwestern.edu' (histonet <@t> lists.utsouthwestern.edu)
Subject: RE: [Histonet] RE: lab math: ihc dilution

You are right I got it wrong, see no matter how many time you run the math, you still come up with problems

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
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liz <@t> premierlab.com
www.premierlab.com

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-----Original Message-----
From: Truscott, Tom [mailto:ttruscot <@t> vetmed.wsu.edu]
Sent: Friday, November 21, 2014 1:01 PM
To: Kathleen Jones; Sheryl Stephenson; histonet <@t> lists.utsouthwestern.edu; Elizabeth Chlipala
Subject: RE: [Histonet] RE: lab math: ihc dilution

When I solve your equation V1 X 1560 ug/ml = 10ml X 0.8 ug/ml I get .005 ml. Therefore the dilution would be closer to 1:2000. I would usually divide the 1560 ug/ml by 0.8 ug/ml and get 1950 therefore 1:1950 would be more accurate . Tom T

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kathleen Jones
Sent: Friday, November 21, 2014 11:50 AM
To: Sheryl Stephenson; histonet <@t> lists.utsouthwestern.edu; Elizabeth Chlipala
Subject: [Histonet] RE: lab math: ihc dilution

Hi Liz, 
 
Wouldn't you dilute in 999.5ul of diluent, not 9999.5. 1ml=1000ul
 
Otherwise I got the same answer...
 
Kathleen
 
Kathleen Jones
Research Technologist
Pathology/Microbiology
AVC - UPEI
(902)213-2207


>>> Elizabeth Chlipala <liz <@t> premierlab.com> 21/11/2014 3:35 PM >>>
Sheryl

Here is how I would approach the problem.  We would use the basic mathematical equation of the following  (in order for this to work you need to keep the units the same)

V1 x C1 = V2 x C2

V1 - volume of stock solution needed - this is what you are solving for
C1 - concentration of stock solution - you know this its 1.56 mg/ml or
1560 ug/ml (there are 1000 ug in one mg)
V2 - volume of final solution needed - I put an arbitrary volume of 10 mls needed
C2 - concentration of final solution - you know this its 0.8ug/ml

V1 x 1560 ug/ml = 10ml x 0.8 ug/ml --- see how I made sure the units were the same I changed the mg/ml to ug/ml

V1 = .0005 mls  and then I would need 9.9995 mls of diluent to make up my 0.8ug/ml concentration, you can change this to be expressed in microliters by multiplying each number by 1000.  

So in uls -  V1 = 0.5 uls and 9999.5 uls

from there you can determine the titer which is your total volume divided by the volume of concentrated antibody

10/.0005

That would be a 1:20,000 dilution

For a dilution such as this I would make up a stock solution of either
1:100 or 1:1000 and then dilute from there.

Now its time for the rest of you to check my math, did I do this correctly?????  I went over it a few times, but you never know..............

Have a GREAT Weekend!

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
liz <@t> premierlab.com
www.premierlab.com

March 10, 2014 is Histotechnology Professionals Day

Ship to Address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Stephenson, Sheryl
Sent: Friday, November 21, 2014 12:00 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] lab math: ihc dilution

To all you math whiz out there, please help with this math dilution.

If my Ab conc is 1.56 mg/mL and they want to optimize to 0.8 ug/mL, what dilution should I use?


Thanks,

Sheryl 




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