Determining Protein Length from Western Blot

[raymanray]

Hello Forum,

please take a look at this image

http://imageshack.us/photo/my-images/98/0bigg.jpg/

It shows a Western blot of an extract of proteins.

My question is how can you determine the length of both proteins A and B graphically?

Could you please help me?

 

[Andy Resnick]

Hello Forum,

please take a look at this image

http://imageshack.us/photo/my-images/98/0bigg.jpg/

It shows a Western blot of an extract of proteins.

My question is how can you determine the length of both proteins A and B graphically?

Could you please help me?

ITYM 'mass' of the protein. IIRC, the mass scale is set using the ladder (the set of spots on the left), and the masses of unknown proteins is then read off the scale.

 

[Ygggdrasil]

The distance migrated by proteins in an SDS-PAGE gel is related to the logarithm of the molecular weight. So, you can measure the distance migrated from the bottom of the well for each of the protein standards in lane 1, plot these distances versus the logarithms of their molecular weights, and find the best-fit line. This best-fit line can then be used to estimate the molecular weights of your samples in lane 2.

 

[raymanray]

Thank you for your answers.

 

[raymanray]

The distance migrated by proteins in an SDS-PAGE gel is related to the logarithm of the molecular weight. So, you can measure the distance migrated from the bottom of the well for each of the protein standards in lane 1, plot these distances versus the logarithms of their molecular weights, and find the best-fit line. This best-fit line can then be used to estimate the molecular weights of your samples in lane 2.

Thank you for your answer.

I read on other websites that I have to measure the distance from the side above from where the proteins started.

How can I plot the distances and their molecular weights? Can you tell me step-by-step please?