What follows is part of protocol for ECM digestion. " The pepsin was first solubilized in 0.1 M HCl and added to the adipose matrix at a ratio of 1 mg pepsin for every 10 mg lyophilized ECM. The adipose matrix was digested for
48 h at room temperature under constant stirring.
👉{{{{Subsequently, the pH was raised to 7.4 using 1 M NaOH and the matrix was diluted to 15 mg ml 1 using 10x PBS so that the final solution contained
1x PBS.}}}}👈
In this protocol I can't figure out the role of PBS with Osmolarity and pH and off course the meaning of 10x and 1x. could you please explain it to me?
Role of pbs is usually to maintain the osmolarity and ph of solution at their required level. The meaning of reaching from 10X to 1X is tenfold dilution of PBS in the solution. E.g 1ml of PBS ( 10X) and 9 ml of diluent will make concentration of PBS to 1X in that solution.
Simpler way of dilution is calculated as:
Required concentration/Given concentration X total volume= volume from stock
Here required is 1X, given is 10X and total volume can be 10ml. 1/10x 10= 1. So 1 ml from 10X stock need to be taken and total volume is makeup by adding 9ml of diluent. (Total volume-volume of stock taken)
Similarly if total volume is 50ml then, 1/10 x50= 5 ( 50-5=45) , 5ml stock and 45ml diluent
If dilution is changed from 10X to 50X, then calculation will be 1/50 x50=1ml ( 50-1=49), 1ml stock and 49ml diluent
Hi,
You can Pick 1 ml of 10X and dilute with 9 ml of distilled water if you desire to have 10 ml of 1X. You can convert it for other volumes.
A.
Role of pbs is usually to maintain the osmolarity and ph of solution at their required level. The meaning of reaching from 10X to 1X is tenfold dilution of PBS in the solution. E.g 1ml of PBS ( 10X) and 9 ml of diluent will make concentration of PBS to 1X in that solution.
Simpler way of dilution is calculated as:
Required concentration/Given concentration X total volume= volume from stock
Here required is 1X, given is 10X and total volume can be 10ml. 1/10x 10= 1. So 1 ml from 10X stock need to be taken and total volume is makeup by adding 9ml of diluent. (Total volume-volume of stock taken)
Similarly if total volume is 50ml then, 1/10 x50= 5 ( 50-5=45) , 5ml stock and 45ml diluent
If dilution is changed from 10X to 50X, then calculation will be 1/50 x50=1ml ( 50-1=49), 1ml stock and 49ml diluent
Mr. Reza, I want to know.. How much your volume in the stock??
Thank u
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