Thermo Scientific™

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

 Related applications:
Anatomical Pathology
Take advantage of a senstive detection system based on biotin-free, polymer technology with the Thermo Scientific™ Lab Vision™ UltraVision™ LP Detection System: HRP Polymer/DAB Plus 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: Diaminobenzidine (DAB)

  • Specificity: anti-Mouse IgG (H+L), anti-Rabbit IgG (H+L)

  • Enzyme: Peroxidase

  • Chromogen/Substrate: Diaminobenzidine (DAB)

Includes:

TL-015-HD: 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 DAB Plus Substrate (TA-015-HSX), 1mL DAB Plus Chromogen (TA-001-HCX)

TL-060-HD: 60mL Hydrogen Peroxide Block (TA-060-HP), 60mL Ultra V Block (TA-060-UB), 60mL Primary Antibody Enhancer (TL-060-PB), 60mL HRP Polymer (TL-060-PH), 60mL DAB Plus Substrate (TA-060-HSX), 2mL DAB Plus Chromogen (TA-002-HCX)

TL-125-HD: 125mL Hydrogen Peroxide Block (TA-125-HP), 125mL Ultra V Block (TA-125-UB), 125mL Primary Antibody Enhancer (TL-125-PB), 125mL HRP Polymer (TL-125-PH), 125mL DAB Plus Substrate (TA-125-HSX), 4mL DAB Plus Chromogen (TA-004-HCX)

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

12. Wash 4 times in buffer.

13. Apply HRP Polymer and incubate for 15 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. Add 1 drop (40μL) DAB Plus Chromogen to 2mL of DAB Plus Substrate, mix by swirling and apply to tissue. Incubate

for 5 minutes, depending on the desired stain intensity.

16. Wash 4 times in DI water.

17. Counterstain and cover slip using a permanent 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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