Author Topic: Confusion about antibody dilution  (Read 9770 times)

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Offline doctor25

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Confusion about antibody dilution
« on: February 15, 2008, 02:52:53 AM »
I have one question concerning antibody dilution .
I got one vial   containing 200 μg IgG in 1.0 ml of PBS from SAnta Cruz Biotechnology,Inc .If they say the working dilution is 1:400,what does it mean
that I take for exemple   1 μl of my stock solution plus 399 of diluent? or   do I take 1:400 as the recommended working antibody concetration and  calculate  the needed solvent quantity?


 Thank you.

Confusion about antibody dilution
« on: February 15, 2008, 02:52:53 AM »

Offline CanuckPhD

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Re: Confusion about antibody dilution
« Reply #1 on: February 15, 2008, 10:34:44 AM »
Yes they mean to dilute 1ul stock solution (200 ug/ml) in 399ul of diluent. This confusion is why it is much better to report the concentration of antibody used as ug/ml NOT the dilution factor. After all it is much easier to repeat an experiment from the literature if know that someone used a dilution of 1ug/ml. If the paper says 1:500, you are left wondering if their stock was 1 mg/ml or 0.1 mg/ml.

It would make all of our lives easier if journals demanded ug/ml instead of dilution factors for publication. Just a pet peeve of mine.

Offline doctor25

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Re: Confusion about antibody dilution
« Reply #2 on: February 17, 2008, 10:28:31 PM »
Thank you very much!I was very confused about this and I think It doesn't make sens because the antibody from different companies has different concentrations.As you said they should express the working dilution of antibody by something like 1mg/l,....
it should be easier to use litterature from other researches
Again thanks!.

Offline ImmunoNYC

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Re: Confusion about antibody dilution
« Reply #3 on: April 15, 2008, 11:00:21 AM »
Yes they mean to dilute 1ul stock solution (200 ug/ml) in 399ul of diluent. This confusion is why it is much better to report the concentration of antibody used as ug/ml NOT the dilution factor. After all it is much easier to repeat an experiment from the literature if know that someone used a dilution of 1ug/ml. If the paper says 1:500, you are left wondering if their stock was 1 mg/ml or 0.1 mg/ml.

It would make all of our lives easier if journals demanded ug/ml instead of dilution factors for publication. Just a pet peeve of mine.
Here here, well said, I couldn't possibly agree with you more!

Offline dutchmaninvienna

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Re: Confusion about antibody dilution
« Reply #4 on: April 16, 2008, 06:55:52 AM »
Yes they mean to dilute 1ul stock solution (200 ug/ml) in 399ul of diluent. This confusion is why it is much better to report the concentration of antibody used as ug/ml NOT the dilution factor. After all it is much easier to repeat an experiment from the literature if know that someone used a dilution of 1ug/ml. If the paper says 1:500, you are left wondering if their stock was 1 mg/ml or 0.1 mg/ml.

It would make all of our lives easier if journals demanded ug/ml instead of dilution factors for publication. Just a pet peeve of mine.
Here here, well said, I couldn't possibly agree with you more!

In principle this is right, on the other hand there are antibodies sold without the precise description of how much antibody is in the solution (for instance CNPase from sternberger inc). In those cases one can only give the dilution. Furthermore although not stated specifically in the articles, if the dilution is given as 1:500, I would assume that the authors give the dilution from the antibody as it came in the bottle. In the end every one still has to find out their own optimal solution so the dilutions suggested by the vendors or authors are only good for finding out the range in which the antibody should be tested.
« Last Edit: April 16, 2008, 06:58:07 AM by dutchmaninvienna »
Jan Bauer, PhD
Associate Professor
Div. of Neuroimmunology
Center for Brain Research
Medical University of Vienna, Austria

Offline eli

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Re: Confusion about antibody dilution
« Reply #5 on: June 15, 2008, 04:20:56 PM »
Well, I do understand all your explanations.
(@doctor25: thanks for asking, that´s just what I was wondering...)
But nevertheless:
HOW do I really know, that 200 µg/1.0 ml doesn´t mean that the antibody is allready prediluted, when calculating the dilution factor which is recommended?  ???

eli

Re: Confusion about antibody dilution
« Reply #5 on: June 15, 2008, 04:20:56 PM »