Collagen Type IV Antibody Staining Protocol for Immunohistochemistry


Description: Collagen IV is comprised of a third chain (alpha-3) together with the 2 classical ones, alpha-1 and alpha-2. COL4A3 gene is localized to 2q36-q37 by analysis of somatic cell hybrids and by in situ hybridization. The alpha-3 chain of type IV collagen induces autoimmune Good pasture syndrome. Mutations in the COL4A4 and COL4A3 genes cause familial benign hematuria.


Primary Antibody

Name: Collagen IV IHC Antibody

Clone: Mouse Monoclonal: Col-30

Supplier: IHC World

Catalog Number: IW-MA1030

Dilution: Ready to Use

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic
Images: Search image

Additional Information:
Species Reactivity: Human, rat
Fixation: Formalin fixed paraffin embedded sections or acetone fixed frozen sections.
Positive Control: Skin, kidney.
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: Normal serum blocking is not needed for this RTU antibody; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary