- Catalog number:
- E-IR-R101
- Applications:
- IHC
- Size:
- 1mL
- Transportation temperature:
- 2~8°C
- Storage:
- 2~8°C
- Leadtime:
- 7~10 days
- Producer manual
Introduction
Elabscience® DAB Detection kit is based on immunohistochemical detection system. Secondary antibody conjugated with HRP which can release the oxygen from substrate H202, then oxygen oxidizes DAB to brown insoluble substance. The site of specific antigen in tissue slices can be observed the brown-yellow color reaction by microscopy.
Instructions
DAB Concentrate (20×) [E-IR-R101A] is concentrated, diluted with DAB Substrate [E-IR-R101B] to 1 × working solution before use.
For example: take 10 μL DAB Concentrate (20×) [E-IR-R101A], add to 190μL DAB Substrate [E-IR-R101B].
Components
Cat. | Products | 1 mL | 3 mL | 6 mL | Storage |
E-IR-R101A | DAB Concentrate (20×) | 1 mL | 3 mL | 6 mL | 2~8°C |
E-IR-R101B | DAB Substrate | 20 mL | 60 mL | 120 mL | 2~8°C |
Manual | One Copy | ||||
Experimental Procedure
1. Dewax and hydrate the paraffin slice
2. Repair antigen of the tissue slice according to special requirements of applied primary antibody.
3. Incubate with 3% H2O2 for 10 min to eliminate endogenous peroxidase activity. Wash with PBS or TBS, 2 min× 3 times.
4. Drain the PBS with absorbent paper, then add Blocking Buffer to the slice. Incubate for 30 min at 37°C.
5. Dry the liquid around the slice with absorbent paper, and draw a circle around the tissue with an oily pen. Add primary antibody with proper dilution ratio, incubate at room temperature or 37°C for 1~2h or at 4°C overnight (then rewarm at 37°C for 30 min). Wash with PBS or TBS, 2 min × 3 times.
6. Add Secondary antibody to the slice (refer to the manual of the secondary antibody). Wash with PBS or TBS, 2 min×3 times.
7. Prepare the DAB working solution Refer to the Instructions above. This working solution must be prepared before use, and store in dark for 4 h, discard the residue working solution.
8. Add appropriate amount of DAB working solution to the slice, incubate for 5~10 min at RT. Take control of the DAB coloration period, the color of tan or brownish yellow is the positive signal. Avoid of excessive reaction.
9. Wash the slice with deionized water to terminate the chromogenic reaction, then operate the procedures of counterstaining, dehydrating, transparentizing and sealing.
Storage
Store at 2~8°C in dark. Avoid of freezing. Valid for 12 months.
The reagents are valid within 6 months after opening.
Cautions
1. For maximal assay performance, this reagent should be used within 12 months. Avoid freeze / thaw cycles.
2. This kit is for research use only. For your safety and health, please wear the lab coat and disposable gloves before the experiments.
3. The positive and negative controls should be set up during the experiment to prevent possible false positive and false negative results.
4. Please dry the liquid around the slice during the experiment to prevent weakening the dyeing intensity.
Introduction
Elabscience® DAB Detection kit is based on immunohistochemical detection system. Secondary antibody conjugated with HRP which can release the oxygen from substrate H202, then oxygen oxidizes DAB to brown insoluble substance. The site of specific antigen in tissue slices can be observed the brown-yellow color reaction by microscopy.
Instructions
DAB Concentrate (20×) [E-IR-R101A] is concentrated, diluted with DAB Substrate [E-IR-R101B] to 1 × working solution before use.
For example: take 10 μL DAB Concentrate (20×) [E-IR-R101A], add to 190μL DAB Substrate [E-IR-R101B].
Components
Cat. | Products | 1 mL | 3 mL | 6 mL | Storage |
E-IR-R101A | DAB Concentrate (20×) | 1 mL | 3 mL | 6 mL | 2~8°C |
E-IR-R101B | DAB Substrate | 20 mL | 60 mL | 120 mL | 2~8°C |
Manual | One Copy | ||||
Experimental Procedure
1. Dewax and hydrate the paraffin slice
2. Repair antigen of the tissue slice according to special requirements of applied primary antibody.
3. Incubate with 3% H2O2 for 10 min to eliminate endogenous peroxidase activity. Wash with PBS or TBS, 2 min× 3 times.
4. Drain the PBS with absorbent paper, then add Blocking Buffer to the slice. Incubate for 30 min at 37°C.
5. Dry the liquid around the slice with absorbent paper, and draw a circle around the tissue with an oily pen. Add primary antibody with proper dilution ratio, incubate at room temperature or 37°C for 1~2h or at 4°C overnight (then rewarm at 37°C for 30 min). Wash with PBS or TBS, 2 min × 3 times.
6. Add Secondary antibody to the slice (refer to the manual of the secondary antibody). Wash with PBS or TBS, 2 min×3 times.
7. Prepare the DAB working solution Refer to the Instructions above. This working solution must be prepared before use, and store in dark for 4 h, discard the residue working solution.
8. Add appropriate amount of DAB working solution to the slice, incubate for 5~10 min at RT. Take control of the DAB coloration period, the color of tan or brownish yellow is the positive signal. Avoid of excessive reaction.
9. Wash the slice with deionized water to terminate the chromogenic reaction, then operate the procedures of counterstaining, dehydrating, transparentizing and sealing.
Storage
Store at 2~8°C in dark. Avoid of freezing. Valid for 12 months.
The reagents are valid within 6 months after opening.
Cautions
1. For maximal assay performance, this reagent should be used within 12 months. Avoid freeze / thaw cycles.
2. This kit is for research use only. For your safety and health, please wear the lab coat and disposable gloves before the experiments.
3. The positive and negative controls should be set up during the experiment to prevent possible false positive and false negative results.
4. Please dry the liquid around the slice during the experiment to prevent weakening the dyeing intensity.
Introduction
Elabscience® DAB Detection kit is based on immunohistochemical detection system. Secondary antibody conjugated with HRP which can release the oxygen from substrate H202, then oxygen oxidizes DAB to brown insoluble substance. The site of specific antigen in tissue slices can be observed the brown-yellow color reaction by microscopy.
Instructions
DAB Concentrate (20×) [E-IR-R101A] is concentrated, diluted with DAB Substrate [E-IR-R101B] to 1 × working solution before use.
For example: take 10 μL DAB Concentrate (20×) [E-IR-R101A], add to 190μL DAB Substrate [E-IR-R101B].
Components
Cat. | Products | 1 mL | 3 mL | 6 mL | Storage |
E-IR-R101A | DAB Concentrate (20×) | 1 mL | 3 mL | 6 mL | 2~8°C |
E-IR-R101B | DAB Substrate | 20 mL | 60 mL | 120 mL | 2~8°C |
Manual | One Copy | ||||
Experimental Procedure
1. Dewax and hydrate the paraffin slice
2. Repair antigen of the tissue slice according to special requirements of applied primary antibody.
3. Incubate with 3% H2O2 for 10 min to eliminate endogenous peroxidase activity. Wash with PBS or TBS, 2 min× 3 times.
4. Drain the PBS with absorbent paper, then add Blocking Buffer to the slice. Incubate for 30 min at 37°C.
5. Dry the liquid around the slice with absorbent paper, and draw a circle around the tissue with an oily pen. Add primary antibody with proper dilution ratio, incubate at room temperature or 37°C for 1~2h or at 4°C overnight (then rewarm at 37°C for 30 min). Wash with PBS or TBS, 2 min × 3 times.
6. Add Secondary antibody to the slice (refer to the manual of the secondary antibody). Wash with PBS or TBS, 2 min×3 times.
7. Prepare the DAB working solution Refer to the Instructions above. This working solution must be prepared before use, and store in dark for 4 h, discard the residue working solution.
8. Add appropriate amount of DAB working solution to the slice, incubate for 5~10 min at RT. Take control of the DAB coloration period, the color of tan or brownish yellow is the positive signal. Avoid of excessive reaction.
9. Wash the slice with deionized water to terminate the chromogenic reaction, then operate the procedures of counterstaining, dehydrating, transparentizing and sealing.
Storage
Store at 2~8°C in dark. Avoid of freezing. Valid for 12 months.
The reagents are valid within 6 months after opening.
Cautions
1. For maximal assay performance, this reagent should be used within 12 months. Avoid freeze / thaw cycles.
2. This kit is for research use only. For your safety and health, please wear the lab coat and disposable gloves before the experiments.
3. The positive and negative controls should be set up during the experiment to prevent possible false positive and false negative results.
4. Please dry the liquid around the slice during the experiment to prevent weakening the dyeing intensity.
Introduction
Elabscience® DAB Detection kit is based on immunohistochemical detection system. Secondary antibody conjugated with HRP which can release the oxygen from substrate H202, then oxygen oxidizes DAB to brown insoluble substance. The site of specific antigen in tissue slices can be observed the brown-yellow color reaction by microscopy.
Instructions
DAB Concentrate (20×) [E-IR-R101A] is concentrated, diluted with DAB Substrate [E-IR-R101B] to 1 × working solution before use.
For example: take 10 μL DAB Concentrate (20×) [E-IR-R101A], add to 190μL DAB Substrate [E-IR-R101B].
Components
Cat. | Products | 1 mL | 3 mL | 6 mL | Storage |
E-IR-R101A | DAB Concentrate (20×) | 1 mL | 3 mL | 6 mL | 2~8°C |
E-IR-R101B | DAB Substrate | 20 mL | 60 mL | 120 mL | 2~8°C |
Manual | One Copy | ||||
Experimental Procedure
1. Dewax and hydrate the paraffin slice
2. Repair antigen of the tissue slice according to special requirements of applied primary antibody.
3. Incubate with 3% H2O2 for 10 min to eliminate endogenous peroxidase activity. Wash with PBS or TBS, 2 min× 3 times.
4. Drain the PBS with absorbent paper, then add Blocking Buffer to the slice. Incubate for 30 min at 37°C.
5. Dry the liquid around the slice with absorbent paper, and draw a circle around the tissue with an oily pen. Add primary antibody with proper dilution ratio, incubate at room temperature or 37°C for 1~2h or at 4°C overnight (then rewarm at 37°C for 30 min). Wash with PBS or TBS, 2 min × 3 times.
6. Add Secondary antibody to the slice (refer to the manual of the secondary antibody). Wash with PBS or TBS, 2 min×3 times.
7. Prepare the DAB working solution Refer to the Instructions above. This working solution must be prepared before use, and store in dark for 4 h, discard the residue working solution.
8. Add appropriate amount of DAB working solution to the slice, incubate for 5~10 min at RT. Take control of the DAB coloration period, the color of tan or brownish yellow is the positive signal. Avoid of excessive reaction.
9. Wash the slice with deionized water to terminate the chromogenic reaction, then operate the procedures of counterstaining, dehydrating, transparentizing and sealing.
Storage
Store at 2~8°C in dark. Avoid of freezing. Valid for 12 months.
The reagents are valid within 6 months after opening.
Cautions
1. For maximal assay performance, this reagent should be used within 12 months. Avoid freeze / thaw cycles.
2. This kit is for research use only. For your safety and health, please wear the lab coat and disposable gloves before the experiments.
3. The positive and negative controls should be set up during the experiment to prevent possible false positive and false negative results.
4. Please dry the liquid around the slice during the experiment to prevent weakening the dyeing intensity.
