Storage And Stability - Leica BIOSYSTEMS BOND Polymer Refine Detection DS9800 Mode D'emploi

BOND Polymer Refine Detection
Catalog No: DS9800
Intended Use
This detection system is for in vitro diagnostic use.
BOND Polymer Refine Detection is a biotin-free, polymeric horseradish peroxidase (HRP)-linker antibody conjugate system for the
detection of tissue-bound mouse and rabbit IgG and some mouse IgM primary antibodies. It is intended for staining sections of formalin-
fixed, paraffin-embedded tissue on the BOND automated system (includes Leica BOND-MAX system and Leica BOND-III system).
The clinical interpretation of any staining or its absence should be complemented by morphological studies and proper controls.
They should be evaluated within the context of the patient's clinical history and other diagnostic tests by a qualified pathologist.
The BOND Polymer Refine Detection Kit must be used with laboratory best practice in the use of tissue controls. For assurance,
laboratories should stain each patient sample in conjunction with positive, negative, and other tissue specific controls as needed.
Summary and Explanation
Immunohistochemical techniques can be used to demonstrate the presence of antigens in tissue and cells (see "Using BOND Reagents"
in your BOND user documentation).
BOND Polymer Refine Detection utilizes a novel controlled polymerization technology to prepare polymeric HRP-linker antibody
conjugates. The detection system avoids the use of streptavidin and biotin, and therefore eliminates non-specific staining as a result of
endogenous biotin.
BOND Polymer Refine Detection works as follows:
• The specimen is incubated with hydrogen peroxide to quench endogenous peroxidase activity.
• A user-supplied specific primary antibody is applied.
• Post Primary IgG linker reagent localizes mouse antibodies.
• Poly-HRP IgG reagent localizes rabbit antibodies.
• The substrate chromogen, 3,3'-Diaminobenzidine tetrahydrochloride hydrate (DAB), visualizes the complex via a brown precipitate.
• Hematoxylin (blue) counterstaining allows the visualization of cell nuclei.
Using BOND Polymer Refine Detection in combination with the BOND automated system reduces the possibility of human error and
inherent variability resulting from individual reagent dilution, manual pipetting and reagent application.
Reagents Provided
Reagents sufficient for 200–300 tests
1. Peroxide Block (30 mL) 3–4% (v/v) Hydrogen peroxide.
2. Post Primary (30 mL) Rabbit anti mouse IgG (<10 μg/mL) in 10% (v/v) animal serum in tris-buffered saline/0.1% ProClin™ 950.
3. Polymer (30 mL) Anti-rabbit Poly-HRP-IgG (<25μg/mL) containing 10% (v/v) animal serum in tris-buffered saline/0.1% ProClin™ 950.
4. DAB Part 1 (2.4 mL) 66 mM 3,3'-Diaminobenzidine tetrahydrochloride hydrate, in a stabilizer solution.
5. DAB Part B (30 mL) ≤0.1% (v/v) Hydrogen Peroxide in a stabilizer solution.
6. DAB Part B (30 mL) ≤0.1% (v/v) Hydrogen Peroxide in a stabilizer solution.
7. Hematoxylin (30 mL) <0.1% Hematoxylin.
Dilution and Mixing
BOND Polymer Refine Detection is optimized for use on the BOND system. Reconstitution, mixing, dilution, or titration of these reagents
is not required.
Materials Required But Not Provided
Refer to "Using BOND Reagents" in your BOND user documentation for a complete list of materials required for specimen treatment and
immunohistochemical staining using the BOND system.

Storage and Stability

Store at 2–8 °C. Do not freeze. Do not use after the expiration date indicated on the tray handle label. Return to 2–8 °C immediately after
use.
There are no obvious signs to indicate instability of this product, therefore positive and negative controls should be run simultaneously
with unknown specimens (refer to "Quality Control" in the "Using BOND Reagents" section of your BOND user documentation).
If unexpected staining is observed that cannot be explained by variations in laboratory procedures, and a problem with the detection
system is suspected, contact your local distributor or the regional office of Leica Biosystems immediately.
Storage conditions other than those specified above must be verified by the user .
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