Biology Reference
In-Depth Information
Ca calibration
For entry of pCa
K
Ca
calculation (see A32..G47)
Solution conditions
Regression analysis (see I29-K35)
7.2
pH
B=
5.12535712 Intermed
4.954 [EGTA]tot (mM)
0.15
M ionic equiv (0.5*sum |zi|Ci)
Gamma H=
0.76295887 H activity coefficient
0.9980 r^2
23
C
Log [H] =
−
7.0825011
Range for linear regression for scatchard
5
mM EGTA
[H] (M) =
8.27E
−
08
should be linear electrode/scatchard slope
500
ml bottle
K
Ca
Assn=
6.363E + 06
Log
K
Ca
= 6.80365
6.86839 = log
K
Ca
from scatchard
K
d
=
1.57E-07
M or
0.1572
m
M
K
Ca
Discn M=
1.572E
−
07
1.35E
−
07 =
K
Ca
dissociation from scatchard
" (nM)=
157.2
135.4 nM
Initial
Ca-free
Ca-total
ml 100 mM
V-Ca
Ca-free
Ca-free
Ca-bound
B/F
Regresn
Inter
Recalculated
CaCl
2
pCa
(nM)
(mM)
(mV)
(M)
(nM)
(mM)
line B/F
mediate
(nM)
pCa
1
8.5000
3.162
0.099
0.493
−
152
5.38E
−
09
5.383
0.099
18319.405
35857.379
−
4.86E
−
03
2.75
8.561
2
8.0000
10
0.299
1.496
−
142.6
1.14E
−
08
11.355
0.299
26341.654
34376.672
−
4.65E
−
03
8.70
8.060
3
7.5000
31.623
0.838
4.188
−
131.5
2.74E
−
08
27.411
0.838
30553.384
30400.143
−
4.12E
−
03
27.55
7.560
4
7.0000
100
1.944
9.722
−
117.1
8.60E
−
08
85.997
1.944
22608.359
22226.140
−
3.01E
−
03
87.48
7.058
5
6.5000
316.228
3.340
16.701
−
101.4
2.99E
−
07
299.131
3.340
11165.474
11917.990
−
1.61E
−
03
280.25
6.552
6
6.0000
1000
4.322
21.609
−
85
1.10E
−
06
1099.966
4.321
3928.010
4674.610
−
6.32E
−
04
924.58
6.034
7
5.5000
3162.278
4.766
23.831
−
70.4
3.51E
−
06
3506.127
4.763
1358.415
1409.423
−
1.87E
−
04
3381.53
5.471
8
5.0000
10000
4.932
24.662
−
51.9
1.52E
−
05
15232.059
4.917
322.822
268.545
−
2.13E
−
05
17,312.44
4.762
9
4.5000
31622.777
5.007
25.034
−
38.1
4.56E
−
05
45563.879
4.961
108.884
−
55.893
5.30E
−
05
63,647.15
4.196
10
3.0000
1,000,000
5.999
29.995
0
9.38E
−
04
938455.736
5.061
5.392
−
790.218
1.05E
−
03
1,046,090.19
2.980
11
3.0000
1,000,000
5.999
29.995
0
9.38E
−
04
938455.736
5.061
5.392
−
790.218
1.05E
−
03
1,046,090.19
2.980
12
3.0000
1,000,000
5.999
29.995
0
9.38E
−
04
938455.736
5.061
5.392
−
790.218
1.05E
−
03
1,046,090.19
2.980
From regression
2.0
10 mM
10 mM
28.2
3.0
1mM
1mM
0.8
[
EGTA]tot
K-Ca-EGTA
4.0
100
M
100
M
29.6
4.953608052 mM
7.38567143
10^6/M
m
m
−
×
Avg slope=
28.9
99.07% % pure
6.868389983 = log K
Slope (mV)=
29
Regression I/J14-: 21
Regression I/J13-: 21
mV offset at 1 mM Ca=
0.8
Slope
B/F intercept
Slope
B/F intercept
-
7385.67143
36585.72
−
6516.46
32752.75
Temperature and ionic strength correction
SE of coeff
135.41
548.75
435.32
1663.88
Std cond
Ionic str
Final
Final
R
^2
0.9980
571.40
0.9697
2324.14
Temp
20
incl T eff
23
C
F
stat
2,975
6
224
7
I-Eq
0.100
0.150
0.150
M
Delta H
Valence
Reg sum Sq
9.71E + 08
1.96E + 06
1.21E + 09
3.78E + 07
Stoich
Log
K
Log
K
Log
K
K
(M)
kcal/mol
2*x*y
Scatchard plot
Electrode calibration
const
K1
40
4.E + 04
9.47
9.3576
9.3138
2.060E + 09
−
5.8
8
K2
8.85
8.7657
8.7219
5.271E + 08
−
5.8
6
0
K3
2.66
2.6038
2.6038
4.016E + 02
0
4
3.E + 04
9
8
7
6
5
4
3
2
K4
2
1.9719
1.9719
9.374E + 01
0
2
−
40
Data
Regression
KCa
10.97
10.7453
10.6840
4.831E + 10
−
8.1
16
2.E + 04
KCa2
5.3
5.1315
5.1315
1.353E + 05
0
12
−
80
Log f
0.109225
0.12327007
1.E + 04
Temp
293
296
−
120
Initial pCa
Recalculated
No EGTA
A
0.507424
0.51006648
0.E + 00
Epsil
80.1057
79.0197311
−
160
0.0
2.0
4.0
6.0
200
−
−
1.E + 04
pCa
Bound (mM)
Fig. 6
Excel spreadsheet used to prepare Ca
2
þ
calibration bu
V
ers using a Ca
2
þ
electrode. This version is used when you want to start
with the pCa of the calibration solutions as input and determine how much total Ca
2
þ
is needed to achieve the desired free [Ca
2
þ
]. It also
allows updating of the apparent K
0
Ca
and free [Ca
2
þ
] in the calibration solutions. This and related spreadsheets can be freely downloaded
(see text for details).
