Thermo Scientific™

Lab Vision™ UltraVision™ LP Detection System: HRP Polymer/AEC Chromogen

 Related applications:
Anatomical Pathology
Employ a senstive detection system based on biotin-free, polymer technology with the Thermo Scientific™ Lab Vision™ UltraVision™ LP Detection System: HRP Polymer/AEC Chromogen.

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Description

UltraVision LP is the latest technology in polymeric labeling. Polymer detection methods have been shown to provide increased sensitivity and detection simplicity. This second-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 LP, you use less antibody and obtain better signal-to-noise ratios. UltraVision LP 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: Peroxidase

  • Chromogen/Substrate: 3-Amino-9-ethylcarbazole (AEC)

Includes:

TL-015-HA: 15mL Hydrogen Peroxide Block (TA-015-HP), 15mL Ultra V Block (TA-015-UB), 15mL Primary Antibody Enhancer (TL-015-PB), 15mL HRP Polymer (TL-015-PH), 15mL AEC Single Solution (TA-015-SA)

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.

WARNING: Reproductive Harm - www.P65Warnings.ca.gov
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. 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. 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 Primary Antibody Enhancer and incubate for 20 minutes at room temperature.

12. Wash 4 times in buffer.

13. Apply 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.)

14. Wash 4 times in buffer.

15. Apply ready-to-use AEC Single Solution to tissue. Incubate for 10 minutes. Modify incubation time to optimize staining in your laboratory.

16. Wash 4 times in DI water.

17. 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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