PAI-1 / Serpin E ELISA Kit
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- Target
- PAI-1 / Serpin E
- Reactivity
- Human
- Detection Method
- Colorimetric
- Method Type
- Sandwich ELISA
- Detection Range
- 156-10000 pg/mL
- Minimum Detection Limit
- 156 pg/mL
- Application
- ELISA
- Analytical Method
- Quantitative
- Sensitivity
- < 10 pg/mL
- Components
- 1. One 96-well plate pre-coated with anti-human PAI-1 antibody 2. Lyophilized human PAI-1 standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-human PAI-1 antibody (Concentrated): 130 µl.
- Material not included
- 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L
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- Comment
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This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-PAI-1 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-PAI-1 polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the PAI-1 amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of PAI-1 can be calculated.
- Plate
- Pre-coated
- Reagent Preparation
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- Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.
- Sample Preparation
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Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Tissue lysate and cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with heparin or EDTA as the anticoagulant. Centrifuge for 15 min at 1000 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Citrate can not be used as anticoagulant here. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP. - Restrictions
- For Research Use only
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- Preservative
- Sodium azide, Thimerosal (Merthiolate)
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- Target
- PAI-1 / Serpin E
- Background
- Plasminogen activator inhibitor-1 (PAI-1) also known as endothelial plasminogen activator inhibitor or serpin E1 is a protein that in humans is encoded by the SERPINE1 gene. This gene is approximately 12.2 kb long and contains 9 exons, and located on the seventh chromosome (7q21.3-q22). PAI-1 is a member of the serine protease inhibitor family that inhibits tissue-type plasminogen activator and urokinase-type plasminogen activator. PAI-1 is mainly produced by the endothelium (cells lining blood vessels), but is also secreted by other tissue types, such as adipose tissue. PAI-1 is present in increased levels in various disease states (such as a number of forms of cancer), as well as in obesity and the metabolic syndrome. It has been linked to the increased occurrence of thrombosis in patients with these conditions.
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