Thermo Scientific™

Lab Vision™ UltraVision™ ONE Detection System: HRP Polymer (Ready-To-Use)

 Related applications:
Anatomical Pathology
Take advantage of a one-step polymer system providing increased sensitivity, time savings and detection simplicity with the Thermo Scientific™ Lab Vision™ UltraVision™ ONE Detection System: HRP Polymer (Ready-To-Use).

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Description

UltraVision ONE is a robust ONE-step polymer system that provides increased sensitivity, time savings and detection simplicity. The UltraVision ONE HRP polymer is an innovative, patented technology. It consists of smaller amino acid polymer subunits that minimize conflicts in binding the target protein. Decreased binding conflicts result in more consistent staining and better signal amplification. Ultimately, this gives the user higher sensitivity and antibody efficiency. UltraVision ONE HRP polymer allows the use of less antibody to obtain better signal-to-noise ratios. This system is also biotin-free, which eliminates background staining found with traditional biotin-based detection methods. For optimal interpretation of results, appropriate positive and negative controls must be included.
  • Specificity: anti-Mouse IgG (H+L), anti-Rabbit IgG (H+L)

  • Enzyme: Peroxidase

  • Chromogen/Substrate: None provided

Includes:

TL-060-HL: 60mL Ultra V Block (TA-060-UB), 60mL UltraVision ONE HRP Polymer (TL-060-PHJ)

TL-125-HL: 125mL Ultra V Block (TA-125-UB), 125mL UltraVision ONE HRP Polymer (TL-125-PHJ)

Note:

UltraVision ONE HRP polymer detection system detects rabbit and mouse immunoglobulins bound to an antigen in tissue sections. The specific primary antibody is located by a universal secondary antibody polymer formulation. The amino acid polymer is conjugated to horseradish peroxidase and the Fab fragments of goat anti-rabbit and goat anti-mouse. The polymer complex is then visualized with an appropriate substrate/chromogen.

Staining Protocol (kit components in bold):

NOTE: The appropriate controls, especially negative controls, must be included with every manual or automated slide run. The inclusion of negative controls will aid in accurate interpretation of the staining results and help in determining false positives. Refer to the warnings and precaution section for details.

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. To reduce nonspecific background staining due to endogenous peroxidase, incubate slide in Hydrogen Peroxide Block for 10-15 minutes.

6. Wash 4 times in buffer.

7. Optional: 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.)

8. Wash (Optional).

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

10. Wash 4 times in buffer.

11. Apply UltraVision ONE HRP Polymer and incubate for 30 minutes at room temperature. (NOTE: HRP Polymer is light sensitive. Please avoid unnecessary light exposure and store in opaque vial.)

12. Wash 4 times in buffer.

13. Incubate with peroxidase-compatible chromogen of choice according to manufacturer's recommendations. Modify incubation time to optimize staining in your laboratory.

14. Wash 4 times in DI water.

15. Counterstain and cover slip using an 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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