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 Thermo Scientific™ Lab Vision UltraVision Detection System: anti-Polyvalent, AP/Fast Red (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: Alkaline Phosphatase

  • Chromogen/Substrate: Fast-Red/Naphthol Phosphate Buffer

Includes:

TP-015-AF: 15mL Ultra V Block (TA-015-UB), 15mL Biotinylated Goat Anti-Polyvalent (TP-015-BN), 15mL Streptavidin Alkaline Phosphatase (TS-015-AP), 15mL Naphthol Phosphate Substrate (TA-015-AFS), three Fast-Red Tablets (TA-003-AFC)

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. Wash 2 times in buffer.

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

4. Wash 4 times in buffer.

5. (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.

6. Rinse (Optional).

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

8. Wash 4 times in buffer.

9. Apply Biotinylated Goat Anti-Polyvalent and incubate for 10 minutes at room temperature. (Some users have experienced better staining when incubating for 20 minutes)

10. Wash 4 times in buffer.

11. Apply Streptavidin Alkaline Phosphatase and incubate for 10 minutes at room temperature. (Some users have experienced better staining when incubating for 20 minutes)

12. Rinse 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.

WARNING: Handle with care and dispose of according to all regulations.

14. Counterstain and cover slip.

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



Documents

Manuals & protocols