Elastin Antibody Staining Protocol for Immunohistochemistry


Description: Elastin is a polymeric protein found in connective tissue that imparts the property of elasticity to vertebrate elastic tissue. It is synthesized and secreted as a soluble, single-chain protein (tropoelastin) which undergoes a number of post-ribosomal modifications prior to its organization into an elastic fiber in the extracellular space. Once secreted, tropoelastin molecules are joined covalently via chemical modification and cross-linking of specific lysyl residues to form the mature insoluble elastin. Ultrastructurally, it is predominantly an amorphous material which may change its morphology with ageing and different disease states. The abnormal accumulation of elastic tissue in blood vessels is found in atherosclerosis and hypertension. Genetic defects in the elastin molecule may result in heritable diseases such as Marfan's syndrome, pseudoxanthoma elasticum and the Bushke-Ollendorf syndrome.


Primary Antibody

Name: Elastin Antibody

Clone: BA-4, Mouse anti-Human

Supplier: Novocastra

Catalog Number: NCL-Elastin

Dilution: 1:200 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 minutes/room temperature

Antigen Retrieval
Device: Incubator
Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101)
Heat/Cool Temperature: 37 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: Extracellular elastic fibers
Images: Search image

Additional Information:
Tissue Type: Kidney, skin, small intestine, liver.
Fixation: Formalin fixed paraffin sections
Positive Control: Kidney, skin, small intestine, liver.
Negative Control: Omit primary antibody, isotype control or absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary


This antibody does not work on acetone fixed frozen sections.