Thermo Scientific™

Lab Vision™ UltraVision™ Detection System: anti-Polyvalent

 Related applications:
Anatomical Pathology
Employ a versatile and robust biotin-streptavidin detection system for research applications and cost-conscious laboratories with the Thermo Scientific™ Lab Vision™ UltraVision™ Detection System: anti-Polyvalent, HRP/DAB (Ready-To-Use).

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Description

This technique involves the sequential incubation of the specimen with an unconjugated primary antibody specific to the target antigen, a biotinylated secondary antibody that reacts with the primary antibody, enzyme-labeled streptavidin, and substrate/chromogen.
  • Labeled streptavidin-biotin immunoenzymatic antigen detection system
  • Species: Goat

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

  • Enzyme: Peroxidase

  • Chromogen/Substrate: Diaminobenzidine (DAB)

Includes:

TP-015-HD: 15mL Hydrogen Peroxide Block (TA-015-HP), 15mL Ultra V Block (TA-015-UB), 15mL Biotinylated Goat Anti-Polyvalent (TP-015-BN), 15mL Streptavidin Peroxidase (TS-015-HR), 15mL DAB Plus Substrate (TA-015-HSX), 1mL DAB PlusChromogen (TA-001-HCX)

Note:

UltraVision detection system detects a specific antibody bound to an antigen in tissue sections. The specific antibody is located by a biotin-conjugated secondary antibody. This step is followed by the addition of a streptavidin-enzyme conjugate that binds to the biotin present on the secondary antibody. The specific antibody, secondary antibody, and streptavidin-enzyme complex is then visualized with an appropriate substrate/chromogen.

Staining Protocol (kit components in bold):

1. Deparaffinize and rehydrate tissue section.

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

3. Wash 2 times in buffer.

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

5. Wash 4 times in buffer.

6. (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.

7. Rinse (Optional).

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

9. Wash 4 times in buffer.

10. Apply Biotinylated Goat Anti-Polyvalent and incubate for 10 minutes at room temperature.

11. Wash 4 times in buffer.

12. Apply Streptavidin Peroxidase and incubate for 10 minutes at room temperature.

13. Rinse 4 times in buffer.

14. Add 1 drop (40μL) DAB Plus Chromogen to 2mL of DAB Plus Substrate, mix by swirling and apply to tissue. Incubate for 5-15 minutes, depending on the desired stain intensity.

WARNING: DAB is a suspected carcinogen. Handle with care and dispose of according to all regulations.

15. Counterstain and cover slip using a permanent mounting media.

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



Documents

Manuals & protocols