Collagen Induced Arthritis: Reagents and Assays

Staining for Collagen

Staining Kit for Type II Collagen

	View product insert
	Request Quote

Reagents Provided (Catalog CIIST)

1. Monoclonal antibody to type II collagen, biotinylated. 
2. Streptavidin-Peroxidase Concentrate 
3. Blocking Buffer. 
4. Antibody Dilution buffer. 
5. Streptvvidin-Peroxidase Dilution Buffer. 

Reagents not provided:

1. Phosphate Buffered Saline (PBS)
2. Insoluble Chromagen (such as DAB)
3. Mounting media
4. 2.5% Gluteraldehyde
5. 2% Bovine testicular hyaluronidase
6. Hydrogen Peroxidase
7. Methanol
8. Immunohistochemical compatible mounting media

Protocol 

1. Fix tissue sections (or cells) with 2.5% gluteraldehyde in PBS for 30 minutes at 4°C.

   Note: Fixed paraffin embedded tissues can also be used.  Deparaffinize the sections with water and continue with step 2.

2. Wash sections 3 times with PBS and gently shake off excess.

3. Treat the sections with 2% bovine testicular hyaluronidase dissolved in PBS, pH 7.4, for 30 minutes at 25°C. 

4. Inactivate endogenous peroxidase by incubating the sections with 1% hydrogen peroxide in methanol for 30 minutes. 

5. Incubate tissue sections with Blocking Buffer for 30 minutes at room temperature.

6. Dilute monoclonal antibody to working concentration with Antibody Dilution Buffer. 

7. Incubate tissues sections with the diluted monoclonal antibody for 1 hour at room temperature.

8. Wash sections 3 times with PBS and gently shake off excess.

9. Dilute Strept-HRP (100 ul) in 10 ml Strept-HRP Diluent. 

10. Incubate tissue sections or cells for 1 hour at room temperature.

11. Wash sections 3 times with PBS and gently shake off excess.

12. Develop sections with an insoluble chromagen such as DAB (brown color).

13. Coverslip with an immunohistochemical compatible mounting media.

 

Product inserts are for information use only.