How to draw graphs for SDS PAGE results?

last updated by

canalon 12 years, 9 months ago

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- February 23, 2012 at 3:47 pm #16128
  craterblizz
  Participant
  Ive got one gel with molecular weight marker: Phosphorylase B, BSA, Ovalbumin, Carbonic anhydrase, Lactoglobulin and Aprotinin.
  Then ive got 4 gels with same protein of 150 Kda giving two bands of 50 KDa and 25 KDa. is there anyway i can draw graphs from this data?
- February 23, 2012 at 6:25 pm #109839
  JackBean
  Participant
  and what kind of graph do you want to make?
  How is it that 150 KDa protein gives 50 and 25 KDa?
  The only I can think of is calibration to determine exact size of your proteins.
- February 23, 2012 at 7:27 pm #109840
  EnricoPallazzo
  Participant
  Looking for a calibration curve?
  On the X-axis plot the migration distance of each band (in mm)
  On the Y-axis plot the log of the molecular masses of the proteins
  Then you`ll get a (hopefully) linear slope from which you can calculate the exact masses of the unknown proteins: http://chemistry.ewu.edu/jcorkill/biochem/PAGESDS.gif
  quote JackBean:
  How is it that 150 KDa protein gives 50 and 25 KDa?
  –> IgG
- February 24, 2012 at 2:53 am #109843
  craterblizz
  Participant
  yes that is IgG (150 KDa) split into light and heavy chain fragment, hence 50 KDa and 25 KDa.
  By migration distance you mean Rf value? dist travelled by the band/ distance travelled by the dye right? If yes, then the X axis value shouldn’t be in ‘mm’ right?
  Also, is there anyway i can find out the standard Rf values for the molecular markers i mentioned?
- February 24, 2012 at 2:59 am #109845
  canalon
  Participant
  If your protein ladder was bought from any reputable company, it is likely to be available on their website. Just look the item number on their catalogue. Or give them a phone call.
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