Fibrinogen（FITC） Product Manual
Fibrinogen(FITC) Instruction Manual
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[Product Name]
Fibrinogen(FITC)
[Packaging Specifications]
0.2ml/vial(FITC).
[Intended Use]
Immunohistochemical staining is performed in addition to routine staining to provide adjunctive information for diagnosis.
[Principle]
Immunohistochemistry (IHC) uses the specific binding of antigen and antibody. A chemical reaction develops the label on the antibody (fluorochrome, enzyme, metal ion or isotope) to detect, localize and characterize tissue and cellular antigens. This reagent uses specific binding of the fibrinogen antibody to target-cell protein, and a chemical reaction develops the fluorochrome label for qualitative detection of the antigen, aiding the diagnosis and differential diagnosis of nephritis. Any positive or negative result should be interpreted by a pathologist together with morphology, clinical findings and other tests, and is not a stand-alone diagnostic indicator.
This reagent is an FITC-labeled rabbit polyclonal antibody.
[Main Components]
Composed of Fibrinogen（FITC） antibody and buffer.
The following instruments and materials must be supplied by the user:
[Storage Conditions and Validity Period]
Store at 2-8°C. The shelf life is 18 months.
Take this product out of the 2-8°C refrigerator only when it is to be used, and return it to the 2-8°C refrigerator immediately after use.
The production date and expiry date are shown on the product label.
[Sample Requirements]
1. Applicable specimen type: paraffin-embedded tissue fixed in 10% neutral (phosphate-buffered) formalin.
2. Specimen collection: surgical specimens.
3. Fixation: the interval from sampling to fixation should not exceed 1 hour, and fixation for 6-48 hours is appropriate.
4. Fixative: 10% neutral (phosphate-buffered) formalin.
5. Section thickness: 3-5 μm.
6. Specimen storage: do not store at room temperature (25±2°C) for more than 5 weeks.
[Test Method]
1. Reagent Preparation
This is a concentrated reagent and must be diluted before use. The recommended dilution is 1:60.
2. Specimen Preparation
Sectioning: sections about 3-5 μm thick.
Baking: 60°C for 120 minutes.
Deparaffinization and hydration: xylene (or xylene substitute) for 10 minutes × 2, absolute ethanol for 5 minutes × 2, 95% ethanol for 5 minutes, then soak in PBS for 5 minutes.
Antigen retrieval: enzyme retrieval (37°C for 10-20 minutes). Apply an appropriate amount of enzyme retrieval solution onto the circled tissue section. After retrieval, rinse and soak in PBS.
Shake off and carefully wipe away excess liquid on and around the tissue. Apply 1 drop (50-100 μl) of fibrinogen antibody working solution to cover the tissue, incubate at 37°C in the dark for 60 minutes, shake off the liquid, and wash in PBS for 5 minutes × 3.
Mount with 50% buffered glycerol.
Seal the edge of the coverslip with neutral gum.
Observe under a microscope and save photographs.
Precautions:
(1) If the fluorescence background is high, counterstain with 0.01% Evans blue (prepared in PBS) for 5 seconds before mounting.
(2) Do not pick the tissue up at the edge of the slide.
(3) Place the tissue so that it is completely covered by reagents throughout staining.
3. Result Interpretation
3.1 Positive staining: target cells in the tissue section show yellow-green fluorescence, with no background staining.
3.2 Negative staining: target cells in the tissue section do not show yellow-green fluorescence.
4. Quality Control
4.1 Positive control: a control slide must be run with every assay and processed together with the case to compare the procedure. The control is a section known to be fibrinogen-positive by IHC. Commercial control slides may be purchased, or known control samples or cell lines may be prepared as described above. Interpret the control by the same criteria. If the control does not meet the requirements, the batch cannot be interpreted.
4.2 Blank control: follow the positive-control procedure, but replace the primary antibody with PBS during incubation. There should be no staining.
4.3 Use the HE-stained section to determine whether immunostain-positive cells are the target cells or otherwise meaningful cells.
4.4 Regardless of the number of positive cells or the staining intensity, genuine positive expression with correct localization is helpful for diagnosis.
[Limitations of the Assay]
Specimens unsuitable for this assay include tissue with obvious crush artifact; tissue for which the time from excision to fixation exceeds 1 hour or fixation is shorter than 6 hours; tissue not fixed in 10% neutral (phosphate-buffered) formalin (unless that laboratory has validated another fixative); or unstained slides stored at room temperature for more than 5 weeks. These conditions are not suitable for fibrinogen detection.
This reagent is for in vitro use only. It is intended for qualitative detection of fibrinogen in paraffin-embedded tissue fixed in 10% neutral (phosphate-buffered) formalin, and is not applicable to other specimen types or fixatives. Specimen and section quality affect the result. The result is for clinical reference only. If it disagrees with the clinical findings, a pathologist and clinician should consult; it cannot alone confirm a diagnosis or guide treatment.
[Precautions]
This product is for professional use and for in vitro testing only.
Read this manual carefully before the experiment.
Laboratory management should strictly follow the standards for an immunohistochemistry laboratory. Operators must receive basic professional and skills training, be familiar with IHC procedures, wear a lab coat and disposable gloves during the experiment, and use a positive-displacement pipette.
Perform quality control for every experiment.
Unless otherwise specified, prepare reagents with purified water.
Dispose of toxic or hazardous reagents according to the applicable procedures.
Avoid microbial contamination of the reagent, otherwise non-specific staining will increase.
Take appropriate safety precautions when handling or using any specimen or reagent of biological origin.
Do not use the reagent if it is turbid or contains flocculent precipitate.
Once dispensed, this reagent is for single use. Do not return it to the container.
[References]
[1] Miao XF, et al. Technical analysis of immunofluorescence pathology in 165 pediatric renal-biopsy specimens. Guangxi Med J. 2011, 33(2):174-176.
[Manufacturer]
Myabtech Biological Inc.
Address :Room 1402, Building F (Self-designated C-3), No. 18 Shenzhou Road, Huangpu District, Guangzhou City
Telephone:020-89858157