Expert answer:A membrane prep was assayed for NET (NE transporte

Solved by a verified expert:A membrane prep was assayed
for NET (NE transporter) binding using increasing concentrations of
3H-Nisoxetine in triplicate.

Background
binding for each reaction was identified in duplicate using 10 mM desipramine

mg protein

Sample
tube#
DPM
total
bckgrd
specific
per Rxn
dpm/mg
fmol/mg

1
1
837

2
1329

3
924

0.364

4
189

5
168

2
6
1530

7
1398

8
1737

0.306

9
234

10
249

3
11
5061

12
4668

0.377

13
3660

14
867

15
753

4
16
7470

17
7527

18
7824

0.325

19
771

20
873

5
21
9552

22
8763

0.436

23
8547

24
1290

25
1242

Please
answer the following questions:

What is the
average total dpm for each reaction?

What is the
average “background” dpm for each
reaction?

What is the
average specific dpm for each
reaction?

What is the
specific dpm/mg for each reaction?

calculate
the fmol/mg of receptor for each reaction, using the following information:

Specific Activity of the
3H-Nisoxetine= 87.2 Ci/mmol

There are 2.2 x 10^6 dpm/mCi
[Hint: make the units cancel]

Show your
work for converting dpm/mg to fmol/mg – you can get partial credit even if
the answer is wrong

Graph the
resulting fmol/mg (you can use excel or another graphing program). What is your ballpark estimate of the
Bmax?