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Cat. No.: ABNFR2-100 (for 100T)

Cat. No.: ABNFR2-500 (for 500T)

Description

Alcian Blue & Nuclear Fast Red Staining Kit (pH 2.5) is a reagent kit used for staining tissues containing sulfated substances, such as sulfated mucosubstances, and tissues containing carboxyl groups, such as sialomucins, hyaluronic acid, and epithelial acid mucin, using Alcian Blue. It also provides Nuclear Fast Red for counterstaining. This kit is suitable for both paraffin-embedded and frozen tissue sections of various tissues and can be used for staining blood smears and bone marrow smears.

The staining process with this kit does not require additional acidification steps, making it a simpler and more convenient staining procedure.

Alcian Blue, also known as Alcian Blue 8G, Alizarin Cyanine, or C.I. 74240, is a copper phthalocyanine dye originally used for dyeing textile fibers. Its molecular formula is C56H68N16S4Cl4Cu, with a molecular weight of 1298.86, CAS number 33864-99-2. Alcian Blue is a cationic dye, and its central copper-containing phthalocyanine ring is connected to four thiolurea groups through sulfur ether bonds. The moderately basic nature of thiolurea groups imparts a cationic property to Alcian Blue. The mechanism by which Alcian Blue stains acidic substances is not fully clear, but it is generally believed that the cationic thiolurea groups form insoluble complexes with polyanionic entities in tissues, such as carboxyl groups in acidic mucins, resulting in a blue color.

Alcian Blue is a highly specific dye for visualizing acidic mucinous substances, making it suitable for mucin staining. The dye forms salt bonds with the acidic groups in mucins. By adjusting the pH of the Alcian Blue staining solution, different categories of acidic mucins can be distinguished. For example, at pH 1.0, tissues containing sulfated substances, such as acidic sulfated or strongly sulfated mucosubstances, will stain blue, while tissues containing carboxyl groups will not stain. At pH 2.5, tissues with carboxyl groups, such as sialomucins, hyaluronic acid, and epithelial acid mucin, can be stained blue, although the staining intensity for sulfated substances is weaker compared to pH 1.0. Neutral mucins, like those found in gastric mucosa and the neutral mucin in the Brunner's gland area, do not react with Alcian Blue. Nuclear Fast Red counterstaining can be used to stain nuclei in pink or red. The colors of acidic mucinous substances and cell nuclei after Alcian Blue-Nuclear Fast Red staining (pH 2.5) are summarized in the table below:

This staining kit exhibits high sensitivity, low background, and clear and distinct coloring.

Storage

Store at room temperature, protected from light, and valid for one year. For long-term storage without use, it can be stored at 4°C or -20°C.

Precautions

• It is recommended to conduct a preliminary experiment with 1-2 samples when using this kit for the first time to determine optimal staining conditions.
• The staining process should be conducted in a humid chamber to prevent the drying of the staining solution.
• The process of Nuclear Fast Red counterstaining is recommended to be lightly stained. Usually, distinguishing cell nuclei is sufficient; excessively dark staining may affect cell observation.
• Select the Alcian Blue-Nuclear Fast Red staining kit at pH 1.0 or pH 2.5 based on different tissue types. The staining time may also vary for different tissues.
• The Alcian Blue staining solution in this kit is already prepared in an acidic solution, and no additional acidification is required.
• Do not use the Alcian Blue staining solution if precipitation occurs.
• If the Nuclear Fast Red staining solution precipitates when stored in low-temperature conditions, such as in a refrigerator, it can be dissolved by heating in a water bath at 25-50°C. Ensure thorough dissolution and homogenization before use.
• This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, food, or drugs. It should not be stored in regular residential areas.
• For your safety and health, wear laboratory clothing and disposable gloves during operation.

Only for research and not intended for treatment of humans or animals

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