Blood-Brain Barrier Permeability Assay Kit

Cat. No.: B1506000

Storage

Protected from light,Room temperature

Shipped In

Normal

Application

Cell Staining

Overview

The core of the Blood-Brain Barrier (BBB) Permeability Assay Kit lies in using spectrophotometry to detect the concentration of substances that have penetrated the simulated blood-brain barrier. It features simple operation and low cost, making it suitable for rapid screening. No fluorescent labeling or radioactive probes are required; instead, it relies on the inherent light absorption properties of the substances themselves (or supporting chromogenic reagents). It has a low operational threshold and uses widely available equipment (a standard laboratory spectrophotometer is sufficient).

B1506000	Components	50T	Storage
B1506000A	Standard Substance A	2 mL	RT. Store in the dark.
B1506000B	Standard Substance B	2 mL	RT. Store in the dark.
B1506000C	Standard Substance C	2 mL	RT. Store in the dark.
B1506000D	Standard Substance D	2 mL	RT. Store in the dark.
B1506000E	Standard Substance E	2 mL	RT. Store in the dark.
B1506000F	Staining Solution	10 mL	RT
B1506000G	Isolation Solution	50 mL	RT
B1506000H	PBS	1 L	RT

Self-Prepared Consumables and Reagents:

Syringe

Tissue Homogenizer

Operating Steps (for reference only):

a. Take the treated experimental animals (using mice as an example). Intravenously inject 1–200 μl of staining solution (No. F). Within a few seconds to 1 minute, the eyes and skin of the mice will turn blue. Sacrifice the mice after 0.5–1 hour and collect the target brain tissue.

b. Place the brain tissue in a 1.5 ml centrifuge tube, add 1 ml of PBS, quickly homogenize the brain tissue using a tissue homogenizer, and centrifuge at 1000×g for 10 minutes at 4°C.

c. Take the supernatant, add an equal volume of separation solution (No. G), mix well, and incubate at 4°C for 18–24 hours.

d. Centrifuge at 2000×g for 15 minutes at 4°C.

e. Take an appropriate amount of the above solution and dilute it 10-fold. Take 1 ml of the diluted solution and measure the absorbance (OD value) at 620 nm using a spectrophotometer or microplate reader. Meanwhile, determine the OD values of standards A–E, plot a standard curve, and calculate the content of the staining solution in the test samples based on the standard curve.

Measured Values by the Company's Spectrophotometer:

B1506000	Standard Substance A	Standard Substance B	Standard Substance C	Standard Substance D	Standard Substance E
Concentration	39.062mM	9.766mM	2.441mM	0.610mM	0.152mM
OD Value	1.036	0.631	0.234	0.116	0.087

(Only relative changes in permeability can be observed, absolute quantification is not possible)

Precautions:

1. In the blood-brain barrier permeability experiment, the injection volume of the staining solution should be adjusted according to the type of animal and the animal's weight.

2. A low-temperature refrigerated centrifuge should be used for centrifugation.

3. For your safety and health, please wear a lab coat and disposable gloves during operation.

Specifications

Stability And Storage

Store at RT long term (12 months). Store in the dark.

Storage

Protected from light, Room temperature

Shipped In

Normal

Chemical and Physical Properties

• Sensitivity: Light-sensitive