PRotein marker can give concentration of my protein in gel? - (May/16/2007 )

I want to have a rought idea that how much protein in my band of SDSGEL. Does anyone have used this kind of protein marker, giving you the concentration of the band in the total protein marker. by comparing the band in my sample and certain band ( already know the concentration) in the protein marker. I could estimate concentration of my band in my total sample?

sounds bit confusing. anyway, that is what I mean.

Thanks

---

in principle with colloid CBB staining, BUT: how do you know what of the stained bands in the crude mix is your protein of interest ?

using immunoblot technique may not be fully quantitative but may give a useful estimation

---

For protein concentration, I doubt you can use it to quantify your protein.

Bearer is right, blotting might be the way.

Hmm Lowry or Bradford will be a better estimation for quantifying the concentration.

---

I think that it have no sense!
If your protein highly purified ( checked on PAAG) so determine concentration by BCA ( 30 min ) or LOWRY, Bradford or Abs at 280 if you know extinction coeff at this wavelength.
If you have a mixture so determine total protein conc by these methods then make SDSPAAG , scan and determine percent of your targeted protein by calculating band intensities in Image programs like IMAGE or Totallab or others. This simple option included in many programs. Then only multiply percent/100 and total protein mix concentration and determine targeted protein concentration in mixture.

---

By doing the western, I already know which one is my interest in SDSPAGE. and it is from a purification, the size I am sure. But it is very small amount. and very less sample. could be ng sample. That is why i want to see roughly how much on gel, but not bradford or so on.


thanks any more suggestion?

Cathy

---