deutsch- Pancreatic elastase 1
- Tumor M2-PK EDTA plasma
- Tumor M2-PK colorectal cancer screening
- Antibodies
- Quick-Prep
- Brainostic
- Veterinary diagnostic
ScheBo® • Brainostic™ GFAP ELISA Kit
GFAP-ELISA for the quantitative detection of the Specified Risk Material (SRM) of brain and spinal cord in meat products and from swabs
ScheBo® • Biotech AG offers the ScheBo® • Brainostic™ GFAP-ELISA test to detect specified risk material for BSE - brain and spinal cord - in meat products, independent of the species. This patented procedure is highly specific and sensitive. It allows the detection of central nervous system tissue in all meat products and sausages (raw, cooked or requiring boiling in water), as well as on contaminated surfaces and carcasses. This test enables an effective control of the SRM-prohibition and enhances preventive consumer protection with respect to exposure to agents causing BSE and other TSEs. One test is suitable for 42 samples in duplicate, independent of the species. No special safety regulations for the laboratory equipment are necessary.
(0.1 - 1.0% CNS). The average coefficient of variance (CV) was 5.9% (range: 2.6-8.6%).
The inter-assay variance was calculated with four samples (0.1-1.0% CNS), which were tested on ten different days. The mean CV was 9.2% (range: 4.6-13.2 %).
Stability of the packaged samples is determined by their expiry date, Swabs are stable for 2 days at room temperature or for 5 days at 4°C. Storage at -20°C increases their stability. Extracts should be analysed on the day of extraction.
Advantages
- Very sensitive detection of BSE-high risk material
- Highly specific immunochemical detection of CNS by use of monoclonal antibodies
- CNS can be detected in all meat products, heat-treated meat products as well as their homogenized intermediate products and from swabs with one test
- Does not give false positive results caused by cross-reaction with blood
- Species-independent screening for BSE-risk material
Methode
- ELISA
- Detection of CNS by using GFAP (Glial Fibrillary Acidic Protein) as marker protein
- 1 blank, 4 standards, 1 control (each in duplicate)
Basic principle of the assay
The ELISA plate is coated with polyclonal antibodies which only recognize Glial Fibrillary Acidic Protein (GFAP). GFAP from samples of meat products, sausages, contaminated surfaces and standards binds to the antibodies and thus is immobilized on the plate. Then the second antibody conjugated to POD (peroxidase) binds to GFAP during the next incubation. The peroxidase oxidizes TMB (3,3’5,5’-tetra-methyl benzidine), which turns yellow. Finally, the concentration of oxidized TMB is determined photometrically.Assay range
The test kit allows the quantification of GFAP within the range of 0.1% to 1.0%. Values outside this range should be specified as < 0.1% or >1.0% respectively.Precision
The intra-assay variance was evaluated by 10-fold determination of four samples(0.1 - 1.0% CNS). The average coefficient of variance (CV) was 5.9% (range: 2.6-8.6%).
The inter-assay variance was calculated with four samples (0.1-1.0% CNS), which were tested on ten different days. The mean CV was 9.2% (range: 4.6-13.2 %).
Sample material
- Meat products
- Heat-treated meat products as well as their homogenized intermediate products
- Sausages
- Minced meat
- Food containing processed meat
- Swabs
Stability of the packaged samples is determined by their expiry date, Swabs are stable for 2 days at room temperature or for 5 days at 4°C. Storage at -20°C increases their stability. Extracts should be analysed on the day of extraction.
Short protocol for the experienced user
Important: The short protocol is not a substitute for the detailed protocol given in the instruction manual!- Prepare the sample-/washing buffer
- Homogenize and extract samples
- Dilute extract in sample-/washing buffer
- Pipette 50 µl blank, standards (ready-to-use), control (ready-to.use) and diluted samples in duplicate into the ELISA-strips
- Incubate 30 minutes at room temperature
- Wash
- 50 µl anti GFAP-POD (ready-to-use)
- Incubate 15 minutes at room temperature (in the dark)
- Wash
- 100 µl substrate solution (ready-to-use)
- Incubate 5 minutes at room temperature (in the dark)
- Add 100 µl stop solution (ready-to-use)
- Read plate at OD 450 or OD 450 - OD 620
- Evaluate with standard curve using a lin-lin scale
AEO-Certificate
The Main Customs Office of Giessen, Germany issued an AEO-F Certificate (Authorized Economic Operator – Customs Simplifications plus Security and Safety) to ScheBo Biotech AG on September 30, 2011.
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