VZV / HHV-3 Mouse Monoclonal Antibody [Clone ID: Mixed]

CAT#: BM3016

VZV / HHV-3 mouse monoclonal antibody, clone Mixed, Purified

Specifications

Product Data

• Clone Name: Mixed
• Applications: ELISA, IF, IHC, IP, WB
• Recommended Dilution: Suitable for use in ELISA, Immunofluorescence, Immunohistochemistry (Paraffin), Western blot, and Immunoprecipitation.
  This monoclonal antibody mix is intended as an aid for the detection of VZV proteins in cell culture by indirect Immunofluorescent test and in formalin-fixed, paraffin-embedded tissue by biotinylated antibody technique assays.
• Host: Mouse
• Isotype: cocktail
• Clonality: Monoclonal
• Specificity: This product is a mixture of 7 affinity purified Murine antibodies prepared from Tissue Culture fluids of Monoclonal antibody producing hybridoma cells.
  The monoclonals in this mixture are Catalog number BM3009 Clone SG1, Catalog number BM3010 Clone SG1-1, Catalog number BM3011 Clone SG2, Catalog number BM3012 Clone SG3, Catalog number BM3013 Clone SG4, Catalog number BM3014 Clone NCP-1 and Catalog number BM3015 Clone IE-62.
  This monoclonal antibody MIX reacts with precursor-products of VZV glycoprotein I (VZV gE), fully glycosylated VZV gE, Glycoprotein II (VZV gB), glycoprotein III (VZV gH), glycoprotein IV (VZV gI), VZV nucleocapsid protein (NCP) and VZV immediate early protein encoded by VZV gene 62 (IE62). The apparent size of precursor-products of VZV proteins range from 45 kDa to 175 kDa; VZV gE, 82 to 95 kDa; VZV gB, 60 to 140 kDa; VZV gH, 100 to 115 kDa; VZV gI, 45 to 60 kDa; VZV NCP, 155 kDa; VZV IE62, 175 kDa.
  Due to the presence of Fc portion of VZV IgG molecules, this reagent may also bind to structural protein A of Staphylococcus aureus or other microorganisms which may be present in specimens tested by this reagent.
• Formulation: Contains an equal quantity of all 7 Monoclonals in 20mM Sodium Phophate, pH 9.0
  State: Purified
  State: Liquid purified Ig fraction
  Preservative: None
• Concentration: lot specific
• Purification: Protein G Chromatography
• Storage: Upon receipt, store undiluted (in aliquots) at -20°C.
  Avoid repeated freezing and thawing.
• Stability: Shelf life: one year from despatch.
• Background: Varicella Zoster Virus (VZV), a member of the human herpes virus family, causes two distinct clinical manifestations: childhood chickenpox(Varicella) and shingles (zoster). Varicella is the outcome of the primary infection with VZV, whereas, zoster is the result of VZV reactivation from latently infected sensory ganglia which occurs predominantly in aging and immunosuppressed individuals.
  VZV is closely related to the herpes simplex viruses (HSV), sharing much genome homology. The known envelope glycoproteins (gB, gC, gE, gH, gI, gK, gL) correspond with those in HSV, however there is no equivalent of HSV gD.
  VZV virons are spherical and 150-200 nm in diameter. Its lipid envelope encloses the nucleocapsid of 162 capsomeres arranged in a hexagonal form. Its DNA is a single linear, double strand molecule, 125,000 nt long.
• Synonyms: Varizella zoster, HHV3
• Note: Protocol: Immunohistochemistry – Formalin-Fixed, Paraffin-embedded Tissue.
  1. Deparaffinize tissue sections using established procedures.
  2. Hydrate tissues with wash buffer containing 1X phosphate buffered saline (PBS) and 0.1% Tween-20.
  3. Pretreat tissues with 1.7 mg/ml Pronase in 0.05M Tris-HCl, pH 7.5 for 10 minutes.
  4. Stop enzyme reaction by washing tissues three times (3 minutes each) with 0.2% glycine in 1X PBS.
  5. Wash tissue three times (3 minutes each) with wash buffer.
  6. Apply blocking reagent (1X PBS, 5% normal goat serum, 2% BSA (fraction V), 0.1% Tween-20, 0.04% sodium azide) for 7 minutes.
  7. Add MAb to VZV at a dilution of 1:100 in antibody diluent (1% BSA, 0.03% Tween-20) and incubate at room temperature for 30 minutes. Note: the optimal antibody dilution must be determined by each laboratory for their specific application.
  8. Wash tissues three times (3 minutes each) with wash buffer.
  9. Apply secondary antibody (biotinylated goat anti-mouse IgG) diluted in antibody diluent and incubate at room temperature for 30 minutes.
  Note: The optimal antibody dilution must be determined by each laboratory for their specific application.
  10. Quench endogenous peroxidase by addition of 0.6% Hydrogen peroxide in 1X PBS and incubate at room temperature for 8 minutes.
  11. Wash tissues 3 times (3 minutes each) with wash buffer.
  12. Apply Vector Elite ABC-peroxidase according to the manufacturer’s instructions (Vector Laboratories) and incubate at room temperature for 30 minutes.
  13. Apply DAB/NiCl2 three times for a total 15 minutes at room temperature.
  14. Wash tissue slides in deionized water three times.
  15. Counterstain tissue slides with 0.1% acridine orange, 0.05% Safranin O in 1X PBS for 2 minutes.
  16. Wash the slides with water for 5 minutes.
  17. Quickly dehydrate, mount and observe under a light microscope.
  
  Interpretation:
  1. Formalin-fixed paraffin-embedded tissue specimens that are positive for VZV proteins will exhibit a brown-black precipitate at antigen-antibody binding. This sharply contrasts with the red-orange color of the Acridin Orange/Safranin O counterstain. In lesions of VZV-infected skin, the antibody produces marked staining of cell membrane, some cytoplasmic staining and, in some cells, dark nuclear staining. Negative tissue specimens may also display a light red-orange background staining due to the staining of collagen in tissue specimens.