problems with ACK/RBC lysis buffer again

Dear all,

I got several responses from this post. They are very helpful to me.
What I guess is that the molar conc. for KHCO3 is 10 mM not 1.0 mM in
the so-called Bible. It seems the  person who copied it might miss a
dot. Because 1) the amount from one company who sells it use gram unit
(it doesn't tell the molar concentration) is 1.0 g/L, and it works fine
for many years.  2) Also in my previous labs they also use the gram unit
to prepare this buffer. It seems there is no problem by now.  I find
this "error" is because this is the first time I have to prepare this
buffer by myself.


Also I try to use this buffer to lysis RBC in cell suspension from
mouse spleen not human.

Once again thanks to all who reply for my post,

Alex  

attached mail follows:

Dear all,

Does anyone use ACK/RBC lysis buffer to remove RBC in the whole blood
or in the spleen suspension? I get a recipe from a well-know "Bible" and
the following is its components (in 1 Liter):
1) NH4Cl      8.29 g   (or 0.15 M)
2) KHCO3     1.0  g    (or 1.0 mM)
3) Na2-EDTA  37.2 mg (or 0.1 mM)

The FW for each reagent:
   NH4Cl----53.49
   KHCO3----100 
   Na2-EDTA---372.22

Before I prepare the solution I do some math using molar concentration.
And here is my result (in 1 L):


1) NH4Cl      8.0235 g	 (or 0.15 M)
2) KHCO3     0.1  g    (or 1.0 mM)
3) Na2-EDTA  37.2 mg (or 0.1 mM)


To my surprise there is a big difference in the KHCO3.	I checked some
papers in several journals. They just show the molar concentrations but 
not gram in their RBC components, which are 0.15 M for NH4Cl, 1.0 mM for
KHCO3, and 0.1mM for Na2-EDTA. I checked their citations which direct me
to the company who sells this product.What I find is that it just lists
the amount of each reagent in gram unit, which is 8.29 G/L for NH4Cl,
1.0 G/L for KHCO3, and 37.2 mg/L for Na2-EDTA. I am confused: which one
is correct? I know sometimes the second person who copies it for
publication might make a mistake. I wonder if somebody in this group may
be able to find out the original paper	or share your experiences with
me?

Thank you very much for the help in advance,

Alex