Steel fences can come in various styles, including picket fences, privacy fences, and ornamental fences. They can be customized with different designs and finishes to match the aesthetic of a property.
A tubular picket fence is a type of fencing that is made up of vertical pickets or posts that are connected by horizontal rails. The pickets are typically made from tubular steel or aluminum, which makes them durable and resistant to rust and corrosion. The fence is designed to provide a secure and attractive barrier around a property, while also allowing for visibility and airflow.
Steel Fence,Steel Security Fences,wrought iron fencing,Tubular Picket Fencing HEBEI DOUDOU METAL FENCE PRODUCTS CO.,LTD. , https://www.security-fencing.com
Human colon cancer antigen (CCA) elisa kit instruction manual
**Human Colon Cancer Antigen (CCA) ELISA Kit – User Manual**
This ELISA kit is designed for the quantitative determination of Human Colon Cancer Antigen (CCA) in human serum, plasma, urine, cell culture supernatant, and tissue homogenates. The kit employs a sandwich ELISA method, using specific antibodies to capture and detect CCA. The detection is based on an enzyme-linked colorimetric reaction, with absorbance measured at 450 nm.
**Kit Specifications:**
- **Configuration:** 48-well or 96-well plate
- **Standard Dilution:** 1.5 ml × 1 vial
- **Enzyme Standard Reagent:** 3 ml × 1 vial (48-well) / 6 ml × 1 vial (96-well)
- **Storage Conditions:** 2–8°C
- **Shelf Life:** 6 months from the date of manufacture
**Kit Components:**
- Sealing Film: 2 pieces (48-well) / 2 pieces (96-well)
- Standard: 2700 ng/L, 0.5 ml × 1 vial
- Enzyme-labeled Antibody: 3 ml × 1 vial (48-well) / 6 ml × 1 vial (96-well)
- TMB Substrate A & B: 3 ml × 1 vial each
- Wash Buffer: 3 ml × 1 vial (48-well) / 6 ml × 1 vial (96-well)
- Concentrated Wash Solution: 20 ml × 20 times (48-well) / 20 ml × 30 times (96-well)
**Principle of Operation:**
The microplate is pre-coated with anti-CCA monoclonal antibody. After adding the sample and standard, CCA binds to the immobilized antibody. HRP-conjugated secondary antibody then binds to the captured antigen, forming a complex. After washing, TMB substrate is added, and the color develops in proportion to the amount of CCA present. The reaction is stopped with an acidic solution, and the absorbance is read at 450 nm. A standard curve is plotted using known concentrations, and the sample concentration is calculated accordingly.
**Sample Preparation Guidelines:**
- **Serum/Plasma:** Centrifuge at 2000–3000 rpm for 20 minutes after clotting or anticoagulant addition.
- **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge to remove debris. For intracellular components, lyse cells by repeated freezing and thawing before centrifugation.
- **Tissue Homogenate:** Weigh the tissue, add PBS, homogenize, and centrifuge. Store the supernatant at 2–8°C or -20°C.
**Procedure Summary:**
1. Prepare standards and samples with appropriate dilutions.
2. Add 40 μl of sample diluent and 10 μl of sample to each well (final 5× dilution).
3. Incubate at 37°C for 30 minutes.
4. Wash 5 times with diluted wash buffer.
5. Add 50 μl of enzyme-labeled reagent to each well.
6. Incubate again at 37°C for 30 minutes.
7. Add 50 μl of TMB A and B, incubate at 37°C for 15 minutes.
8. Stop the reaction with 50 μl stop solution.
9. Read OD at 450 nm within 15 minutes.
**Notes:**
- Allow the kit to equilibrate at room temperature for 15–30 minutes before use.
- Avoid repeated freeze-thaw cycles.
- Do not mix reagents from different batches.
- Keep all reagents away from light.
- Handle all waste as biohazardous material.
- Ensure accurate pipetting and calibration.
- Always run a standard curve and duplicate wells for better accuracy.
**Performance Characteristics:**
- Intra-assay CV < 9%, Inter-assay CV < 11%
- Correlation coefficient (R²) ≥ 0.95
- Detection range: 0.2 IU/L – 6 IU/L
**Technical Support:**
Free technical support available during working hours. Sample testing services are also offered upon request.
**Storage & Stability:**
Store all components at 2–8°C. Do not freeze the microplate.
**Safety:**
This kit is for research use only. Avoid contact with skin and eyes. Use gloves and lab coat when handling reagents.
**Warranty:**
If any issues arise, please contact our customer service team for assistance.
This manual provides detailed instructions for accurate and reliable CCA detection. Always follow the protocol carefully to ensure optimal results.