DAB-Stained Sample Preservation Solution (For Plant) - BioReagent,for microscopy,Biological Stain

Cat. No.: D1511471

GRADE & PURITY BioReagent, Biological Stain, for Microscopy

Synonyms

Plant Section Preservation Solution (Post-DAB Staining)

Storage

Room temperature

Shipped In

Normal

Overview

  Plant tissues produce various reactive oxygen species (ROS) under stress conditions. Due to their high reactivity and poor stability, ROS are typically analyzed by detecting their downstream products. Hydrogen peroxide is a common ROS. Catalyzed by catalase, it can rapidly react with DAB (3,3'-Diaminobenzidine tetrahydrochloride) to produce a brownish-red product, enabling the localized detection of hydrogen peroxide in tissues. This method is also known as the DAB staining method and is suitable for in situ staining of hydrogen peroxide in living plant tissues.

  The DAB Staining Sample Preservation Solution (for Plant) is specifically designed for preserving plant samples after localized hydrogen peroxide staining. It allows stained samples to be stored stably at room temperature for one week and is primarily suitable for whole-mount staining samples of materials such as young root tips and leaves. Areas with hydrogen peroxide accumulation appear brown to dark brown after staining. This product is for research use only and not intended for clinical diagnosis or other non-research purposes.

Materials to Be Prepared by User

• Fresh plant leaves or roots, tap water, distilled water, 95% ethanol
• Ultrasonic cleaner, magnetic stirrer, electronic balance, filter paper, camera
• DAB Staining Solution (for Plant Use)

Procedure (For Reference Only)

1. Sample Preparation: Collect stressed (e.g., by heavy metals) plant seedlings or root tips. Rinse briefly with tap water and place on filter paper to remove excess moisture.

2. Staining: Immerse the plant seedlings or root tips in the DAB Staining Solution. Stain at room temperature, protected from light, for 2~6 hours, until positive areas develop a dark brown color, while other areas remain nearly colorless or retain the plant's natural color. (Adjust staining time based on plant tenderness and degree of color development.)

3. Destaining (Chlorophyll Removal): Carefully remove the seedlings or leaves with forceps. Rinse by immersing and agitating in distilled water 3~5 times. Place on filter paper to remove excess water, then immerse in 95% ethanol and treat at 40°C for 3~16 hours. The purpose is to remove the chlorophyll from the seedlings or leaves. Replace with fresh 95% ethanol several times during this process.

4. Observation and Preservation: Remove the seedlings or leaves with forceps. Rinse by immersing and agitating in distilled water 3~5 times. Place on filter paper to remove excess water. Transfer the sample to an adequate amount of DAB Sample Preservation Solution and soak for 30 minutes. Subsequently, the sample can be removed for photography. The sample can be stored in this preservation solution at room temperature for up to one week.

Notes

1. Once prepared, the DAB staining working solution should be stored at 4°C, protected from light, and used within one week. Prolonged storage may affect color development.

2. Since hydrogen peroxide is prone to decomposition and any external factors may stimulate its stress-induced production in plants, plant samples should be freshly collected and stained as soon as possible. It is recommended to include negative and positive control groups.

3. Photograph the stained samples as soon as possible after staining to preserve the results.

4. DAB may be carcinogenic. Handle with care and avoid direct contact.

5. For the staining and destaining steps, the following alternative procedure is also recommended: Place the tissue in the staining solution, apply vacuum to -0.1 MPa, maintain the negative pressure for 20~30 minutes, then let it stain at room temperature for 60 minutes. Discard the staining solution. Add 95% ethanol and destain in a water bath at 70~80°C, changing the 95% ethanol every 10 minutes. Destaining can be stopped once the green color of the sample has completely faded.

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

7. Use the reagents as soon as possible after opening to avoid affecting subsequent experimental results.

8. Recommended DAB Staining Solution Product Numbers:

   • D1509200 DAB Staining Solution (1.0 mg/mL, pH 3.8, for Plant Use)

   • D1509202 DAB Staining Solution (1.0 mg/mL, pH 5.5, for Plant Use)

   • D1509203 DAB Staining Solution (2.0 mg/mL, pH 3.8, for Plant Use)

Specifications

Synonyms

Plant Section Preservation Solution (Post-DAB Staining)

Specifications & Purity

BioReagent, for microscopy, Biological Stain

Stability And Storage

Store at room temperature long term (12 months).

Storage

Room temperature

Shipped In

Normal

Grade

Biological Stain, BioReagent, for Microscopy