Volume and Concentration of Reagents and Proteins
● Reagent to use with hemoglobin
○ Deionized/distilled water
○ Two protocols from unrelated sources suggest suspension of hemoglobin in
deionized water
● Concentration of hemoglobin
○ Soluble at 20 mg/mL
○ Recommended concentration for reconstitution, 20 mg/mL
■ From two protocols from unrelated sources
● Volume of combined hemoglobin and water for device (in well)
○ Volume: 250uL
■ Selected based on recommended working volume of Corning for 96 well
plate (75-200uL), but also considering volumes large enough for device to
handle
○ Diameter of well: 7mm
■ Selected based on diameter of standard 96 well plate commonly used in
fluorescent plate readers
○ Height of well: 6.5mm
■ Calculated using equation:
● V= pi*r^2*h
● h=V/pi*r^2
● Next Steps
○ Obtain hemoglobin
○ Design wells to allow for filling of protein and reagent
● Sources
○ http://www.sigmaaldrich.com/content/dam/sigmaaldrich/docs/Sigma/Product_Information_Sheet/2/h7379pis.pdf
○ http://www.sigmaaldrich.com/Graphics/COfAInfo/SigmaSAPQM/SPEC/H7/H7379
/H7379-BULK________SIGMA____.pdf
○ http://www.mpbio.com/product.php?pid=0855914&country=223