# Making 1ug/ml EGF - (Jan/06/2009 )

Hello Everyone

I'm kind of a new student working in lab. And I need to use a 1ug/ml final concentration of EGF for my treatments

I have just purchased the EGF and have no idea how to go about making this...Sorry this is kind of a dumb question...

The instruction said to add sufficient 10mM of 1M HCl to get a peptide concentration of atleast .1mg/ml but they recommend 1 mg/ml.

If I follow 1mg/ml that would mean that I would have to add 1ml of 10mM HCl????

But then I would have to dilute to get to 1ug/ml????

Thank you.

QUOTE (theundergrad09 @ Jan 6 2009, 03:56 PM)
Hello Everyone

I'm kind of a new student working in lab. And I need to use a 1ug/ml final concentration of EGF for my treatments

I have just purchased the EGF and have no idea how to go about making this...Sorry this is kind of a dumb question...

The instruction said to add sufficient 10mM of 1M HCl to get a peptide concentration of atleast .1mg/ml but they recommend 1 mg/ml.

If I follow 1mg/ml that would mean that I would have to add 1ml of 10mM HCl????

But then I would have to dilute to get to 1ug/ml????

Thank you.

It depends on the weight of the EGF in the vial you bought. I recently just got 0.2 mg of EGF from Sigma. If I want a 1 mg/ml solution, I would add 0.2 ml of 10 mM acetic acid (what they recommended for reconstitution). Then, I would need to calculate how much of this working stock to add to my media, which depends on the final volume of treatment solution I need to make (what and how are you going to treat?). Using (C1)(V1) = (C2)(V2):

What you HAVE:(C1=working stock)(V1=unknown) What you WANT: (C2)(V2=100 ml, for example)
(1 mg/ml)(X) = (1 ug/ml)(100 ml)
or
(1 mg/ml)(X) = (0.001 mg/ml)(100 ml)

Then X=0.1 ml.

So, add 0.1 ml (100 ul) of your 1 mg/ml working stock to your medium for treatment.
In my case, I only made 200 ul of my working stock, so I would have 100 ul of my 1 mg/ml stock leftover that I could aliquot and freeze for later.

Does that sound right? Could someone check my math?

-bertie-

Thank you so much! that makes sense!

QUOTE (bertie @ Jan 6 2009, 01:57 PM)
QUOTE (theundergrad09 @ Jan 6 2009, 03:56 PM)
Hello Everyone

I'm kind of a new student working in lab. And I need to use a 1ug/ml final concentration of EGF for my treatments

I have just purchased the EGF and have no idea how to go about making this...Sorry this is kind of a dumb question...

The instruction said to add sufficient 10mM of 1M HCl to get a peptide concentration of atleast .1mg/ml but they recommend 1 mg/ml.

If I follow 1mg/ml that would mean that I would have to add 1ml of 10mM HCl????

But then I would have to dilute to get to 1ug/ml????

Thank you.

It depends on the weight of the EGF in the vial you bought. I recently just got 0.2 mg of EGF from Sigma. If I want a 1 mg/ml solution, I would add 0.2 ml of 10 mM acetic acid (what they recommended for reconstitution). Then, I would need to calculate how much of this working stock to add to my media, which depends on the final volume of treatment solution I need to make (what and how are you going to treat?). Using (C1)(V1) = (C2)(V2):

What you HAVE:(C1=working stock)(V1=unknown) What you WANT: (C2)(V2=100 ul, for example)
(1 mg/ml)(X) = (1 ug/ml)(100 ml)
or
(1 mg/ml)(X) = (0.001 mg/ml)(100 ml)

Then X=0.1 ml.

So, add 0.1 ml (100 ul) of your 1 mg/ml working stock to your medium for treatment.
In my case, I only made 200 ul of my working stock, so I would have 100 ul of my 1 mg/ml stock leftover that I could aliquot and freeze for later.

Does that sound right? Could someone check my math?