help with antibody titrations

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raves
raves's picture
help with antibody titrations

Can someone help me with an urgent problem i have. iam wanting to titre some antibodies to work out the best concentration to use. The stock concentration is 0.2mg/ml. im wanting to do a dilution of my antibodies from 10ug to 5ug to 2ug to 1ug to 0.1ug .
how is the best way to do this when i need approx 200ul volume for each tube

raves

mchinmoyee
mchinmoyee's picture
To find the right

To find the right concentration or titre of antibody to use I prefer to use the concentration as provided by the suppliers for 1st time or can refer if other people have used the same antibody with the same system and can then little fiddle with next time low or high , you will get to know.Mostly as they write dilutions like 1:500-1000 and according to the concentration given in the specification sheet mostly in mg/ml as in your case 0.2mg/ml = 0.2ug/ul so if you use 1:2000 dilution you will be using 0.1ug, 1:10000 is 1ug and so on.

Or

The other way is DOT Blot with different concentration of antibody seperately in one go .

Good Luck

Guo-qiang Huang
Guo-qiang Huang's picture
Are the 10ug, 5ug, 2ug ,1ug 0

Are the 10ug, 5ug, 2ug ,1ug 0.1ug per ml ? or absolute value ( in 200ul volume)?

To make "10ug", you should take 50ul from your stock , then either put on 150ul for your 200ul volume or 950 ul for you 1ml volume.

Duplicate another "10ug", make 1 /2, 1/5, 1/10 and 1/100 solution, you can get all the concentration you want.

samm
samm's picture
wrote:

wrote:

The easiest way to do is to keep the following formula in your head:
Amount of stock=[Required concentration/Stock concentration] x final volume.
Amount of diluent=Final volume - amount of stock

for e.g to get a concentration of 10 ug/ml of antibody in 200 ul volume from a stock of 0.2 mg/ml,
Amount of stock = [10/200] x 200 = 10 ul
Amount of diluent/cell suspension etc = 190 ul.

This works for everything: cells, antibodies, molar amts of rgts etc.