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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
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Name: vWF Antibody |
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Clone: Rabbit anti-Human |
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Supplier: Biocare Medical |
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Catalog Number: CP039A |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
References: