von Willebrand Factor (vWF) Antibody Staining Protocol for Immunohistochemistry


Description: von Willebrand Factor (vWF) is a large glycoprotein with a multimeric structure and a molecular mass ranging from 500 kDa up to more than 10,000 kDa, the latter being the largest known for a soluble human plasma protein. The primary product of the von Willebrand Factor gene, located at chromosome 12p13.2, is a 2813 amino acid protein comprising a signal peptide of 22 amino acids, a large propeptide of 741 amino acids, and a mature vWF molecule containing 2050 amino acids. Expression of the von Willebrand Factor gene is tissue specific and confined to endothelial cells and megakaryocytes. vWF is present in plasma, in the Weibel Pallade bodies of endothelial cells, in the alpha-granuel in megakaryocytes and platelets derived from them, as well as in the subendothelial matrix of the vessel wall. Von Willebrand Factor serves as a carrier for Factor VIII in plasma, protecting the circulating coagulation coenzyme from proteolytic degradation.


Primary Antibody

Name: vWF Antibody

Clone: Rabbit anti-Human

Supplier: Biocare Medical

Catalog Number: CP039A

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 min/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: Cytoplasmic (endothelial cell of blood vessels)
Images: Search image

Additional Information:
Species Reactivity: Human, chicken, sheep, pig and swine
Fixation: Formalin fixed paraffin sections
Positive Control: Tonsil, liver, kidney, heart, skin, normal lung and angiosarcoma
Negative Control: Omit primary antibody, isotype control, 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