10 µM (=5.731 mg/L) ATP in HBS

Use ATP aliquots from Lab 6.1

10 µM ATP, Calcium-free

Lab 6.2: 0.75 mL per coverslip x 2 coverslips x 8 groups = 12 mL

Make 15 mL

• 15 µL 10 mM ATP stock
• in 15 mL Ca- HBS (1 µL 10 mM ATP stock per 1 mL HBS.)

8 1.6 mL aliquots for Lab 6.2

1mg/mL Bradykinin stock
(= 1.0602 mM) Sigma B3259 - 1 mg septum bottle
MW = 106.02
Add 1 mL H2O to the 1 mg in the bottle (NO CALCIUM! - some will be used for the calcium experiments in Lab 7.2)

2 µM Bradykinin in HBS

Use bradykinin aliquots from Lab 6.1

2 µM Bradykinin, Calcium-free
(=2.12 µg/mL)

Lab 6.2: 0.75 mL per coverslip x 2 coverslips x 8 groups = 12 mL

~2 µL 1 mg/mL stock per 1 mL working solution

31.8 µL 1 mg/mL stock in 15 mL Ca-free HBS

Make 8 aliquots of 1.6 mL

Make more if necessary. (Aliquots should have been enough for 15 mL?)

Use aliquots from Lab 6.1

HBS needed for rinsing and to make other solutions

(3 rinses + 1 sham dilution) x 4 expts/grp x 1 mL = 16 mL

Aliquot 25 mL per group

300 mL HBS

300 mL Ca-free HBS

Ionomycin stock

Fisher or Invitrogen (Life Technologies) I24222

1 mM in DMSO or EtOH. 1 mg makes 1.34 mL stock.

3 µM Ionomycin in HBS
Dilute stock 1:333 in HBS: 3 µL/1 mL

24 µL in 8 mL, aliquot 800 µL

They only need 750µL, because they do one experiment in HBS, then two in Ca-free HBS.

3 µM Ionomycin in Ca-free HBS
Dilute stock 1:333 in Ca-free HBS: 3 µL/1 mL
45 µL in 15 mL

Takes a long time to thaw. Vortex.

Aliquot 1.55 mL so they can do two 750 µL experiments