[Histonet] RE: lab math: ihc dilution

[Truscott, Tom]

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
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liz <@t> premierlab.com
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-----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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