Biology Forum › Community › General Discussion › reconstituting protein
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- April 25, 2012 at 10:33 am #16396
biology_06erParticipantHi there,
I was sent a protein (100ug) and the data sheet tells me to reconstitute at 100ug/mL in PBS..does this mean I just add 1mL of PBS to the powder form I have hence I’ll have a final conc of 100ug/mL…
what if I had a powder form of 50ug (as you can get this conc as well sent and the data sheet also says to reconstitute it at 100ug/mL..is this even possible..if i used the formula )
c1*v1=c2*v2
50*v1=100*1mL
v1=100/50
v1=2doesn’t make sence to me..to add "2" mL/s
anyone help with both questions above..i think i have the first one correct
Cheers,
b_06er - April 25, 2012 at 10:54 am #110761JackBeanParticipant
yes, no
you cannot use c1V1 = c2V2, since you’re not diluting a solution but solubilizing a solid phase. Thus, you’re missing V1. What you want is m = c.V (in your case is concentration in g/l, not mol/l), thus you have
50 ug = 100 ug/ml . V1 -> V1 = ? - April 29, 2012 at 1:15 am #110812biology_06erParticipant
Hi, thanks for that…so another question following on from it..if I have 100ug/mL of protein and I plan to run an ELISA using it..and to each well I need (1.5 ug/mL with 100 uL/well)
So for example, if I wanted to use 50 wells with 100uL in each well and 1.5ug/mL in each well would I use the formula
C1*V1=C2*V2
100ug/mL*V1=1.5ug/mL*(50*100uL=5000uL)V1=75uL
So I would add 75uL of my 100ug/mL stock to 4925uL of PBS to make 1.5ug/mL…and if I took 100uL of this and added to each well would I be correct?
Thanks,
b__06er - May 4, 2012 at 1:04 pm #110879JackBeanParticipant
yes, but are you sure, it’s supposed to be 100 ul of only your protein?
(BTW it’s always better to make little bit more of your mix, because you may lack some due to the pipetting error)
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