Ready To Use HP IHC Detection Kit #abs957

abs957-5ml

In Stock

364

Application

Primary antibody is from mouse or rabbit

Substrate for color development

DAB (biphenyldiamine)

Reagent composition

Component number

Component name

Specification

Quantity

A

hydrogen peroxide

5ml

1

B

Super blocking solution

5ml

1

C

Primary Antibody Amplifier

5ml

1

D

Enzyme-labeled secondary antibody polymer

5ml

1

E

DAB A liquid

5ml

1

F

DAB B liquid

160ul

1

Not provided but necessary reagents

Primary antibody (from mouse or rabbit), PBS (phosphate buffer)This kit is the latest generation of immunohistochemical secondary antibody non-biotin detection system based on polymer technology, with stronger signal and simpler operation. Compared with the traditional immunohistochemical secondary antibody kit, it has three characteristics: First, due to the uniqueness of the polymer, the polymer molecule formed by the direct combination of the enzyme molecule and the secondary antibody IgG molecule is highly sensitive and effectively reduces The amount of primary antibody used, the primary antibody reagent can be diluted 2-4 times; secondly, because the use of biotin is avoided, non-specific background coloration caused by biotin can be avoided, and clearer color development and cleaner Background; the third is fast and shorter reagent incubation time.

Storage and expiration date

Store at 2-8°C. Each component can be stored for at least 18 months.

Steps:

Note: All steps are performed at room temperature. Adding reagent can be done directly by titration via a bottle or a pipette. One drop = 30-40ul

1. Dewaxing and hydrating tissue sections

2. Wash in PBS 2-3 times for 5 minutes each

3. Pre-treat tissue sections according to the special requirements of the applied primary antibody

4. Wash 2-3 times in PBS for 5 minutes each

5. Add 100ul A solution and incubate for ten minutes to block endogenous peroxidase to reduce non-specific background staining

6. Wash 2-3 times in PBS for 5 minutes each

7. Add 100ul B solution and incubate for five minutes to reduce non-specific staining. Note: This step should not exceed ten minutes; if the primary antibody is diluted in a buffer containing 5% to 10% normal goat serum, this step can be omitted

8. Wash 2-3 times in PBS for 5 minutes each

9. Add primary antibody and incubate at room temperature or 37°C for 20 minutes

10. Wash with PBS 2-3 times for 5 minutes each

11. Add 100ul C solution and incubate for ten minutes

12. Wash with PBS 2-3 times for 5 minutes each

13. Add 100ul D solution and incubate for ten minutes. Note: This solution is sensitive to light, please avoid light

14. Wash with PBS 2-3 times for 5 minutes each

15. Prepare fresh substrate solution: Mix 30ul of F solution and 1ml of E solution, then prepare fresh substrate solution (the substrate solution can be stored for 2 weeks)

16. Add 100ul of fresh substrate solution and incubate for five minutes

17. Rinse with deionized water

18. Counterstaining and sealing