Description
REF:WDA8P0001
INTENDED USE
PA-Enhancer(Linker) is ready to use. PA-Enhancer(Linker) is intended for signal amplification of primary rabbit antibodies in combination with the PA-Immunochromogenic Reagent in formalin-fixed, paraffin-embedded tissue sections.For in vitro diagnostic use only. For professional use only.
SUMMARY
PA-Enhancer(Linker) provides a 2–3 fold signal amplification fo rabbit antibodies when used as supplement of the PA -Immunochromogenic Reagent.
PRINCIPLE
PA-Enhancer(Linker) contains a mouse anti-rabbit polyclonal antibody, when using together with PA-Immunochromogenic Reagent, PA-Enhancer(Linker) could make a bride for the Enhancer and polymer, then make a three step method reagent and magnify the signal.
PRECAUTION
1. This kit is for in vitro diagnostic use only.
2. Do not reuse, expired products may not be used.
3. The kit is to be used by professionals.
4. Insufficient amount of reagents in the experiment may lead to incorrect results.
5. Avoid contact of reagents with eyes, skin, and mucous membranes. Use protective clothing and gloves.
6. If reagents come in contact with sensitive areas, wash with copious amounts of water. Avoid inhalation of reagents.
7. Ensure that the waste container is empty prior to starting a run on the instrument. If this precaution is not taken, the waste container may overflow and the user risks a slip and fall.
8. PA-Enhancer(Linker) contains material of animal origin. As with any product derived from biological sources, proper handling procedures should be used according to local requirements.
9. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin.
10. Unused solution should be disposed of in accordance with all local, regional, national and international regulations.
MATERIALS
Materials Provided
For WDA8P0001(100 Tests)
| Contents | Main components | Quantities |
| Enhancer (Linker) | Enhancer(Linker) contains a mouse anti-rabbit polyclonal antibody and antibody diluent. | 15 mL/bottl e * 1 |
| Instruction for Use | Instruction for Use | 1 |
Materials Required but Not Provided
1. PA-Sample Release Reagent
2. PA-Immunochromogenic Reagent
3. PA-Retrieval Solution(pH9.0)
4. PA-Retrieval Solution(pH6.0)
5. PA-Bluing Reagent
6. PA-Wash Buffer
7. Microscope slides
8. Positive and negative tissue to use as process controls
9. Distilled or de-ionized water
10. Ethanol absolute
11. Xylene or xylene substitutes
12. Permanent mounting medium
13. Cover glass
14. General purpose laboratory equipment
15. Bright field microscope (4-40x objective magnification)
16. 7ml reagent vial (Tagged with RFID)
Equipment Needed
Fully Automated Pathology Staining System (Model No.: PA-3600).
STORAGE AND STABILITY
1. Store at 2~8°C, valid for 18 months
2. Keep away from sunlight, moisture and heat.
3. Freezing and thawing prohibited.
4. Use within 3 months after opening.
5. Tighten the cap and return to 2~8 °C immediately after use.
6. Do not use after expiration date printed on the vial label.
SPECIMEN COLLECTION AND PREPARATION
The specimens may be formalin-fixed, paraffin embedded tissue sections. Fixation time is dependent on fixative and tissue type/thickness. For example, tissue blocks with a thickness of 3~5 mm should be fixed in neutral-buffered formalin for 18~24 hours. The optimal thickness of paraffin-embedded sections is approximately 3 ~ 5 μm. After sectioning, tissues should be mounted on Microscope Slides and then placed in a 65 ± 2 °C calibrated oven for 1 hour.
The sections should be mounted on the slides as flat and wrinkle-free as possible. Wrinkles will have a negative impact on the staining results.
NOTE: The position of the specimens on the microscope slides must suitable for the PA3600 instrument. Please refer to the User Guide for definition of usable staining area of the microscope slide for the specimen.
TEST PROCEDURE
Used in combination with the Fully Automated Pathology Staining System (Model No.: PA-3600), the process of dewaxing to counterstaining is completed by the instrument.
1. Place the prepared slides in a 65 ± 2 °C calibrated oven for 1 hour.
2. Following the operating instructions of the instrument software.
3. Setting up the protocol using the instrument software and printing labels.
4. Loading the labeled slides into the instrument.
5. Placing reagents into the reagent rack and confirming that the reagent type is correct and that the amount of reagents is sufficient to complete the experiment.
6. Start the operation for automatic staining.
7. After staining is completed, remove the sections and rinse with distilled water.
8. Then dehydrate and transparent the slides, finally mount the slides with permanent mounting medium and cover slipped.
For complete information and operating procedure, please refer to PA 3600 Operation Manual.
RESULT INTERPRETATION
The diaminobenzidine containing Substrate Working Solution gives a brown color at the site of the target antigen recognized by the primary antibody. The brown color should be present on the positive control specimen at the expected localization of the target antigen. If nonspecific staining is present, this will be recognized as a rather diffuse, brown staining on the slides treated with the negative control reagent.Nuclei will be stained blue by the hematoxylin counterstain.
QUALITY CONTROL
Positive control should be set for each batch of experiments to monitor the operation process and the quality of reagents;a positive control can be fished on Refer to Quality Assurance for Design Control and Implementation of Immunohistochemistry Assays; Approved Guideline-Second Edition (I/LA28-A2) CLSI. 2011
Positive and negative control tissues (lab-supplied) should be run for each staining procedure. These quality controls are intended to ensure the validity of the staining procedure,including reagents, tissue processing and instrument performance. It is recommended that control tissues be stained on the same slide as the patient tissue.
Positive Control
The positive control should be a tissue with positive biomarker expression. External Positive control materials should be fresh specimens fixed, processed, and embedded as soon as possible in the same manner as the patient sample(s). One positive external tissue control for each set of test conditions should be included in each staining run.If the positive tissue controls fail to demonstrate positive staining, results with the test specimens should be considered invalid.
Negative Control
The negative control should be a tissue or tissue element with no biomarker expression. Use a negative tissue control fixed, processed, and embedded in a manner identical to the patient sample(s) with each staining run to verify the specificity of the IHC primary antibody for demonstration of the target antigen,and to provide an indication of specific background staining (false positive staining).If specific staining (false positive staining) occurs in the negative tissue control, results with the patient specimens should be considered invalid.
Nonspecific Negative Reagent Control
Use a nonspecific negative reagent control in place of the primary antibody with a section of each patient specimen to evaluate nonspecific staining and allow better interpretation of specific staining at the antigen site.The incubation period for the negative reagent control should correspond to that of the primary antibody.
LIMITATIONS OF PROCEDURE
1. Immunohistochemistry is a multi-step process, each step may influence the result, these include, but are not limited to fixation, antigen retrieval method, incubation time,tissue section thickness, detection kit used and interpretation of the staining results.
2. The recommended protocols are based on exclusive use of Wondfo products.
3. The clinical interpretation of any positive or negative staining should be evaluated within the context of clinical presentation, morphology and other histopathological criteria by a qualified pathologist.
4. The clinical interpretation of any positive or negative staining should be complemented by morphological studies using proper positive and negative internal and external controls as well as other diagnostic tests.
PERFORMANCE CHARACTERISTICS
Positive conformance
The positive control was taken, and the immunohistochemical test was carried out according to the instructions of the manufacturer. The results met the requirements that the positive tissue/cell staining result should be positive, the positioning of the positive staining should be accurate, and there should be no background staining or non specific staining.
Negative conformance
Take a negative control according to the manufacturer’s instructions for immunohistochemical tests,the results met the negative tissue/cell staining results of negative, no background staining or non-specific staining requirements.
Blank control conformance
Take a blank control according to the manufacturer’s instructions for immunohistochemical tests, and antibody diluent was used instead of primary antibody working liquid as a blank control. Immunohistochemical tests were conducted according to the instructions of the manufacturer. The results met the requirements of negative, no background and non-specific staining of the infected tissues/cells.
Intra batch precision
Three tissue slices from the same tissue source containing the target antigen were taken and the same batch of products were used for immunohistochemical detection. The results met the requirements of no obvious difference in staining intensity and localization of tissue slices from the same tissue source.
Inter batch precision
Three tissue slices from the same tissue source containing the target antigen were taken and 3 different batches of products were used for immunohistochemical detection at the same time. The results met the requirements that there is no obvious difference in the intensity and location of staining of tissue slices from the same tissue source with different batches of reagents.
BIBLIOGRAPHY
[2] Wu Bingquan, Liu Yanfang. Immunohistochemical Pathological diagnosis (2nd edition) [M]. Science in Beijing Technology Press, 2013.
[3] He Jianfang, Han An ‘an, WU Qiuliang. Practical immunohistochemical pathological diagnosis [M]. Science Press Society, 2018.
[4] JD Bancroft and A Stevens. Theory and Practice of Histological Techniques. 4th Edition. Churchill Livingstone, New York. 1996.
[5] Clinical and laboratory Standards Institute (CLSI). Protection of Laboratory workers from Occupationally Acquired Infections; Approved Guideline-Fourth Edition CLSI document M29-A4 Wayne, PA 2014.
INDEX OF SYMBOL
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Consult instructions for use |  |
Manufactur er |  |
Date of manufacture |
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In vitro diagnostic medical device |  |
Use-by date | |
Catalogue number |
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Temperature limit |  |
Batch code |  |
Contains sufficient for tests |
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Keep away from sunlight |  |
Abbreviation for volume |  |
CE marking |
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Biological risks |  |
Contains biological material of animal origin |  |
Authorized representative in the European Community/ |
| European Union | |||||
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Importer |  |
Unique device identifier |
