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

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

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

 

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