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| Custom Evaluation for Drug BBB-Permeability (Only in Japan) |
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| 1. Custom Evaluation Schedule |
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【assay buffer：DPBS-H 】
| 10×PBS(+/+) |
100 mL |
| d-glucose |
4.5 g |
| HEPES (1M) |
10 mL |
| DW |
890 mL |
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| Total |
1,000 mL (at 37℃) |
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10 x Dulbecco's Phosphate buffered saline (10 x PBS (+/+)
(Sigma: D1283)
d-glucose (Wako Pure Chemical Industories ：041-00595）
HEPES buffer (1M) (Sigma: H0887) |
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Your compound will be diluted into the concentration of 1.0 μM with assay buffer, DPBS-H.
| Caution 1） |
You may change the concentration of your compound for cytotoxicity and mesurement sensitibity. |
| Caution 2） |
When you need to use organic solvent to dissolve your compound, the final concentraion of the solvent should be under 0.5% (V/V). |
| Caution 3） |
Cytotoxicity of your compound can be evaluated by TEER (trans endothelial electrical resistance) after permeability assay. |
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| No |
Sample |
Solution |
Concentration |
Concentration |
Storage temprature |
Others |
| Ex. |
PCC-1 |
DMSO |
1mM |
20μL |
-20℃ |
- |
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| Drug BBB-Permeability Assay |
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For evaluating the permeability of a molecule through the brain endothelial and pericytes layers of BBB Kit ™ in blood-to-brain direction, the molecule is aplied to the upper compartment of the insert.
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| *Depending on types of molecules or sensitivity measurement, the time points are decided. |
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| *The value obtained with only Transwell® membrane is the background data. The backgound data is needed in the calculation of apparent permeability. |
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t=0 |
ｔ=20 |
t=40 |
t=60 |
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luminal |
○
（assay buffer） |
- |
- |
○ |
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abluminal |
○
（assay buffer） |
○ |
○ |
○ |
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cell + membrane |
− |
- |
- |
○
（if needs） |
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１．Calculation of apparant permeability
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| ２．Evaluation of Papp |
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We obtained a very good correlation between the BBB Kit™ and in vivo permeabilities of drugs. You can evaluate the BBB-permeability by our BBB Kit™, quantitatively.
When you design molecular modifications of your compound or vectors carrying your compound into the brain, you can easily evaluate the BBB-permeability, quantitatively. |
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Papp（×10-6cm/s） |
Permeability |
ex) |
| Permeabilitis |
>20 |
very good |
Antipyrine |
| 10〜20 |
good |
- |
| 2〜10 |
low |
Na-F |
| <2 |
very low |
ESA, sucrose |
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| *Transwell® is trademark of Corning, Incorporated, Corning, NY, USA. |
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MBT-24H
