Biomedical Engineering Reference
In-Depth Information
Arginine hydrochloride 242 (1.15), asparagine 50 (0.38), aspar-
tic acid 20 (0.15), cystine 50 (0.21), glutamine 300 (2.05), glu-
tamic acid 20 (0.14), glutathione 1 (0.003), glycine 10 (0.13),
histidine hydrochloride 18 (0.09), hydroxyproline 20 (0.15),
isoleucine 50 (0.38), leucine 50 (0.38), lysine hydrochloride 40
(0.22), methionine 15 (0.10), phenylalanine 15 (0.09), proline
20 (0.17), serine 30 (0.29), threonine 20 (0.17), tryptophane 5
(0.02), tyrosine 20 (0.11), valine 20 (0.17);
p-Aminobenzoic acid 1 (0.007), biotin 0.2 (0.0008), choline
chloride 3 (0.02), folic acid 1 (0.002), i-inositol 35 (0.19), niaci-
namid 1 (0.008), calcium pantothenate 0.25 (0.0005), pyridox-
ine hydrochloride 1 (0.005), riboflavin 0.2 (0.0005), thiamine
hydrochloride 1 (0.003), vitamin B
12
(cyanocobalamin) 0.005.
-
RPMI 1640 HAT
Supplement 100 ml RPMI 1640 with 1 ml HT stock and 1 ml
A stock.
HT Stock solution
: Make slurry of 408 mg hypoxanthine in
100 ml ddH
2
O, then add 1 M sodium hydroxide until all solid is
dissolved. Dissolve 114 mg thymidine in 100 ml ddH
2
O. Com-
bine both solutions, fill up to 300 ml, and adjust pH with acetic
acid to 10.0. Filter through a 0.2-
µ
m filter and store in aliquots
at −20
◦
C.
A Stock solution
: Vortex 45.4 mg aminopterine (4-aminopteroyl
glutamic acid, sodium salt) in 5 ml ddH
2
O and dissolve the solid
by dropwise addition of 1 M NaOH.Fillupto100ml and adjust
pH to 7.5. Filter through a 0.2-
µ
m filter and store at −20
◦
C.
-
Krebs
-
Henseleit
-
Ringer Buffer
CaCl
2
280 (2.53), KCl 354 (4.74), KH
2
PO
4
162 (1.19), MgSO
4
·
7H
2
O 292 (1.19), NaCl 6923 (118.54), NaHCO
3
2100 (25.00).
-
Ty rode So lut i on
CaCl
2
200 (1.80), KCl 200 (2.68), NaCl 8000 (136.99), NaHCO
3
1000 (11.90), Na
2
HPO4
·
2H
2
O 60 (0.34), Glucose 1000 (5.55).
7.4.3 pH Calibration Buffers
A potassium hydrogentartrate solution, saturated at 25
◦
C
B 0mM potassium hydrogenphthalate: 1.0212 g C
8
H
5
KO
4
per
100 ml
C 5mMphosphate buffer: 0.3403 g KH
2
PO
4
; 0.4450 g Na
2
HPO
4
·
2H
2
O per 100 ml
D .7mM phosphate buffer: 0.1183 g KH
2
PO
4
; 0.5416 g Na
2
HPO
4
·
2H
2
O per 100 ml
E 0mM sodium tetraborate (borax): 0.2012 g Na
2
B
4
O
7
per
100 ml
The pH values derived by the standard buffers are temperature de-
pendent. The pH values between 0 and 60
◦
C aregiveninTable7.5,
which also indicates the change of pH at 1:1 dilution (
∆
pH
1:1
).
