Thermo Scientific™

Lab Vision™ UltraVision™ LPValue Detection System: AP Polymer/Fast Red Chromogen

 Related applications:
Anatomical Pathology
Take advantage of a third-generation polymer system composed of smaller polymer subunits that minimize conflicts in binding the target protein with the Thermo Scientific™ Lab Vision™ UltraVision™ LPValue Detection System: AP Polymer/Fast Red Chromogen.

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Description

UltraVision LPValue is the latest technology in polymeric labeling. Polymer detection methods have been shown to provide increased sensitivity and detection simplicity. This third-generation polymer system is composed of smaller polymer subunits that minimize conflicts in binding the target protein. Decreased binding conflicts result in more consistent staining and better signal amplification.1 Ultimately, this gives the user higher sensitivity and antibody efficiency.2 With UltraVision LPValue, you use less antibody and obtain better signal-to-noise ratios. UltraVision LPValue is also biotin-free, which eliminates background staining found with traditional biotin-based detection methods.
  • Specificity: anti-Mouse IgG (H+L), anti-Rabbit IgG (H+L)

    Enzyme: Alkaline Phosphatase

  • Chromogen/Substrate: Fast Red/Naphthol Phosphate Buffer

Includes:

TL-015-AFS: 15mL Ultra V Block (TA-015-UB), 15mL Value Primary Antibody Enhancer (TL-015-PBS), 15mL Value AP Polymer (TL-015-APS), 15mL Naphthol Phosphate Substrate (TA-015-AFS), 3 Fast Red Tablets (TA-003-AFC)

Note:

UltraVision detection system detects a specific mouse IgG or rabbit IgG antibody bound to an antigen in tissue sections. The specific antibody is located by a universal secondary antibody formulation conjugated to an enzyme-labeled polymer that recognizes mouse and rabbit immunoglobulins. The polymer complex is then visualized with an appropriate substrate/chromogen.

Staining Protocol (kit components in bold):

1. Deparaffinize and rehydrate tissue section.

2. Wash 2 times in buffer.

3. If required, incubate tissue in digestive enzyme (or appropriate pretreatment).

4. Wash 4 times in buffer.

5. Apply Ultra V Block and incubate for 5 minutes at room temperature to block nonspecific background staining. NOTE: Do not exceed 10 minutes or there may be a reduction in desired stain. (May be omitted if primary antibodies are diluted in buffers containing 5-10% normal goat serum.)

6. Wash (Optional).

7. Apply primary antibody and incubate according to manufacturer's recommended protocol.

8. Wash 4 times in buffer.

9. Apply Value Primary Antibody Enhancer and incubate for 20 minutes at room temperature.

10. Wash 4 times in buffer.

11. Incubate with Value AP Polymer anti-Mouse/Rabbit IgG for 30 minutes at room temperature.

12. Wash 4 times in buffer.

13. Add 1 Fast Red tablet to 5mL of Naphthol Phosphate Substrate and shake until tablet is dissolved. Apply to tissue section and incubate for 10-20 minutes or until desired stain intensity is achieved.

14. Wash 4 times in DI water.

15. Counterstain and cover slip using aqueous mounting media.

The specificity and sensitivity of antigen detection is dependent on the specific primary antibody used. General References:

1. Shan-Rong Shi, James Guo, Richard J. Cote, Lillian Young, Debra Hawes, Yan Shi, Sandra Thu, and Clive R. Taylor, Applied Immunohistochemistry & Molecular Morphology, Vol 7, 201-208, 1999.

2. Karen Petrosyan, Rosalba Tamayo, and Daisy Joseph, “Sensitivity of a Novel Biotin-free Detection Reagent (PowerVision+) for Immunohistochemistry” J. Histotechnology, Vol 25, 247-250, 2002.



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