Collagen Immunostaining

Chondrex, Inc. provides immunostaining kits and a series of monoclonal antibodies for staining various types and species of collagen. Our immunostaining kits contain all necessary reagents, such as biotinylated monoclonal or monoclonal antibody cocktail, streptavidin-peroxidase, blocking buffer, and antibody dilution buffer.

Collagen Staining Kits

Monoclonal Antibodies for Immunostaining

Reagents Provided
 

1.  Biotinylated monoclonal antibody, 250 micrograms/ml x 100 microliters

2.  Streptavidin peroxidase, 50 microliters x 2 vials

3.  Blocking buffer, 15 ml

4.  Antibody dilution buffer, 20 ml

5.  Streptavidin peroxidase dilution buffer, 20 ml

Reagents Not Provided

1.  Tissue fixative
 

2.  Phosphate buffered saline (PBS)

3.  2% bovine testicular hyaluronidase

4.  1% hydrogen peroxidase
 

5.  Insoluble chromagen (such as DAB)

6.  Mounting media

Type I Collagen (Human) Staining Kit

Rat monoclonal antibody, 42R (IgM) is highly specific to human type I collagen and does not cross-react with other species of type I collagen, and ideal for staining type I collagen in human tissues.

Type II Collagen (Multi-Species) Staining Kit

The four clone monoclonal antibody cocktail (Arthrogen-CIA®) to type II collagen recognizes the conserved epitopes of various species of type II collagen, such as human, monkey, porcine, rat, mouse, rabbit, equine, and chick, and cross-reacts to all species to type II collagen by more than 95%. Thus, this cocktail can be used for immunoblotting and immunostaining of tissue sections from these species.

Note: Since alpha3 (XI) chain of type XI collagen is identical to alpha1 (II) chain of type II collagen, these monoclonal antibodies cross-react to type XI collagen, but do not cross-react to type I, type III, or type IX collagen.

Type IX Collagen (Multi-Species) Staining Kit

The monoclonal antibody cocktail to type IX collagen recognizes the conserved epitopes on the low molecular weight fragment of human, bovine, rat, mouse, and chick type IX collagen. Thus, this cocktail can be used for immunoblotting and immunostaining of type IX collagen in tissue sections from human, bovine, rat, mouse, and chick.

Note: These anti-type IX monoclonal antibodies do not cross-react to type I, type II, and type XI collagen. The cross-reactivity of type IX in other species (except human, bovine, rat, mouse, and chick) has not been determined.

Figure 1 – Rat External Ear Cartilage 6a) stained by Anti-type IX collagen antibody 6b) stained by anti-type II collagen antibody
 

Type IV Collagen (Glomerular Basement Membrane) Staining

Rat nephritogenic monoclonal antibodies, clone a84 (Catalog # 7061 and 7062) and b35 (Catalog # 7062 and 7064), can be used for immunostaining of rat kidney basement membrane. These monoclonal antibodies specifically recognize the NC1 fragment of rat type IV collagen of glomerular basement membrane. See Rat Glomerular Nephritis.  

Chondrex, Inc. also provides mouse monoclonal antibodies, Clone 3-1-1 (Catalog # 7056, and 7057) and Clone CB243 (Catalog # 7058 and 7059), which recognize the conserved epitopes on the NC1 fragment of mouse and bovine type IV collagen. These monoclonal antibodies are not capable of inducing nephritis in mice, and have been used for immunoblotting.

Figure 2 – Direct immunostaining of cryostat section of rat kidney with monoclonal antibody, Clone b35

A cryostat section of rat kidney was incubated with rat Anti-NC1 of rat type IV collagen monoclonal antibody, Clone b35, and then with FITC-conjugated goat anti-rat IgG antibody.

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