Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

How do I go about making a set of protein standards for protein quantification v


  • Please log in to reply
2 replies to this topic

#1 dette

dette

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 02 June 2014 - 06:49 PM

I have been given a protein of unknown concentration and have been asked to make a set of standards of a protein with a known concentration so we can quantify roughly via dot blot.The aim is to use a floursescent secondary antibody and to visualise via odessy.

 

I need to know how to work out the calculation part. I would like to learn both mass and molar.

 

For example, the information I have for my standard protein is 30mg/ml and 550uM. I want to put for example 1ul dots on the membrane for my blot. I might want several dilutions of my standard, for example ranging from 20pg to 100ng. How do I work it out to get varied concentrations from the stock to my 1ul volumes on my blot?

 

Also, should I do varied dilutions of the unknown protein?

 

Thanks, I'm so new to theis and very confused

 

 

 

 

 

 



#2 neuron

neuron

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 187 posts
5
Neutral

Posted 05 June 2014 - 04:16 AM

30mg/ml or 30ug/ul means you have 30ug in 1ul. Now if you make dilutions like 1:100 and 1:1000, you will get 300 ng and 30 ng respectively. Same way you can dilute it further till you reach to 30pg/ul. Accordingly you can add different dilutions to make standards with keeping volume constant. 



#3 dette

dette

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 08 June 2014 - 08:19 PM

Hey I get it, thank you so much.






Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.