Leica Biosystems Bond Oracle HER2 IHC System - Manual (Page 4)

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Leica Biosystems Bond Oracle HER2 IHC System Instructions for Use TA9145 EN-CE-Rev_E 18/06/2013
Principle of Procedure
The Bond Oracle HER2 IHC System contains components required to complete an
immunohistochemical staining procedure for formalin-fixed, paraffin-embedded tissues.
Following incubation with the ready-to-use HER2 Primary Antibody (clone CB11), this
system employs ready-to-use Compact Polymer technology. The enzymatic conversion of
the subsequently added chromogen results in the formation of a visible reaction product at
the antigenic site. The tissue sections may then be counterstained, dehydrated, cleared and
mounted. Results are interpreted using light microscopy. Control slides with four formalin-fixed,
paraffin-embedded human breast cancer cell lines are provided to validate staining runs. The
four cell lines demonstrate HER2 oncoprotein expression at 0, 1+, 2+ and 3+ intensities. The
staining intensity of these cell lines correlates to both HER2 oncoprotein receptor load per cell
and HER2 gene amplification status.
The Bond Oracle HER2 IHC System (product code TA9145) is for use on the Leica Biosystems’
BOND fully automated, advanced staining system.
Components Provided
The materials listed below (Table 1) are sufficient to stain 150 slides (60 test slides incubated with
HER2 Primary Antibody, 60 corresponding test slides incubated with HER2 Negative Control,
15 HER2 Control Slides incubated with HER2 Primary Antibody and 15 in-house positive tissue
controls incubated with HER2 Primary Antibody). The number of tests is based on the use of
a 150 µL automated dispense per slide. The kit provides materials sufficient for a maximum of
15 individual BOND staining runs.
HER2 Control Slides,
(x15)
Sections of formalin-fixed, paraffin-embedded, human breast cancer
cell lines that demonstrate HER2 oncoprotein expression at 0, 1+, 2+ and
3+ staining intensities when stained in accordance with the protocol
provided. These sections are fully adhered and do not require further
baking.
HER2 Primary
Antibody, 13.5 mL
Contains ready-to-use, affinity-purified, mouse monoclonal IgG antibody,
clone CB11 and 0.35% ProClin
™
950 .
HER2 Negative
Control, 9 mL
Contains ready-to-use mouse IgG at an equivalent concentration to the
HER2 Primary Antibody and 0.35% ProClin
™
950.
Peroxide Block,
22.5 mL
Contains 3-4% hydrogen peroxide.
Post Primary, 22.5 mL
Rabbit anti-mouse IgG (<10 μg/mL) in Tris-buffered saline containing 10%
(v/v) animal serum and 0.09% ProClin
TM
950.
Polymer, 22.5 mL
Poly-HRP goat anti-rabbit IgG (<25 μg/mL) in Tris-buffered saline
containing 10% (v/v) animal serum and 0.09% ProClin
TM
950.
DAB Part 1, 2.25 mL
Contains 66 mM 3,3’-diaminobenzidine tetrahydrochloride, in a stabilizer
solution.
DAB Part B (x2),
22.5 mL
Contains ≤0.1% (v/v) hydrogen peroxide.
Hematoxylin, 22.5 mL Contains <0.1% hematoxylin.
Table 1. Bond Oracle HER2 IHC System components