DAB Peroxidase Substrate Kit (Purple-Blue Color)
Based on 1 publication(s) in Google Scholar
MCE DAB Peroxidase Substrate Kit (Purple-Blue Color) can be used for staining and color development detection in experiments such as immunohistochemistry of cells or tissues, in situ hybridization, Western blotting, and for visualizing endogenous HRP in cells or tissues.
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Storage :
-20℃, 1 year. Keep away from light and avoid repeat freeze-thaw cycles.
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Description & Advantages
3, 3’-Diaminodbenzidine (DAB) is a chromogenic substrate that can be used with horseradish peroxidase (HRP) for immunohistochemistry and immunoblotting applications. Under HRP catalysis, the substrate DAB reacts with hydrogen peroxide, transferring electrons from DAB to the peroxide, producing an insoluble brown product.
MCE DAB Peroxidase Substrate Kit (Purple-Blue Color) has an upgraded formula based on the original, enhancing the sensitivity of color development. The final color will appear gray-blue or purple-blue. This kit can be used for staining and color development detection in experiments such as immunohistochemistry of cells or tissues, in situ hybridization, Western blotting, and for visualizing endogenous HRP in cells or tissues.
Protocol
1. After incubation: Incubate regular tissue sections, cell samples, or membranes with HRP-labeled antibodies or other types of probes, and then wash 3-5 times with the appropriate washing buffer. For tissues or cell samples detecting endogenous HRP, wash them 3-5 times with washing buffer after appropriate fixation, each time for 3-5 min.
2. Prepare DAB working solution: Refer to the table below for preparation (adjust volume as needed).
Note: DAB Concentrate (20×) and DAB Diluent Buffer must be thoroughly mixed to prepare the working color development solution.
3. Incubation with DAB: After washing, remove the washing solution and add the DAB working solution to fully cover the sample. Incubate at room temperature in the dark for 1-30 min or longer.
Note: 1) If no background appears, incubation can continue until the desired color intensity is reached.
2) In Western blot experiments, ensure there is enough DAB working solution for the membrane to move freely.
4. Stop the reaction: Remove the DAB working solution and wash several times with ddH2O to stop the color development reaction.
5. Post-staining and mounting: For cells or tissue samples observed under a microscope, optional counterstaining can be performed, followed by mounting in an aqueous medium. Alternatively, samples can be dehydrated and mounted in an organic mounting medium. For Western blot experiments, wash the membrane with ddH2O, air dry, and store at room temperature.
Publications
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Storage
-20℃, 1 year.
Keep away from light and avoid repeat freeze-thaw cycles.
Attention
1. As Sodium azide (NaN3) inhibits HRP, the reaction system must not contain NaN3.
2. DAB is harmful to humans; handle it carefully, and use proper protective measures to avoid direct contact or inhalation.
3. The working solution should be freshly prepared and used immediately. Freshly prepared DAB solution is yellow-brown, turns gray within minutes, and may precipitate if left too long.
4. Adjust the color development time according to the actual conditions to avoid insufficient or excessive staining.
5. This product is for R&D use only, not for drug, household, or other uses.
6. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K1107-500 T | HY-K1107-3000 T |
|---|---|---|
| DAB Concentrate (20×) | 1.25 mL × 2 | 15 mL |
| DAB Diluent Buffer | 47.5 mL | 142.5 mL × 2 |
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