Nori Canine CD14 ELISA Kit
$461.00 – $832.00
This ELISA kit is for quantification of CD14 in dog. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for CD14 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CD14 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for CD14 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of CD14 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for CD14: Cluster of differentiation 14
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Canine CD14 ELISA Kit Summary
Alternative names for CD14: Cluster of differentiation 14,
Alternative names for canine: dog
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number | A0A1S7J0A8 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 18 pg/mL |
| Detection Range | 93.75-6000 pg/mL |
| Specificity | Canine CD14 |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:
CD14 (cluster of differentiation 14) is a component of the innate immune system. CD14 exists in two forms, one anchored to the membrane by a glycosylphosphatidylinositol tail (mCD14), the other a soluble form (sCD14). Soluble CD14 either appears after shedding of mCD14 (48 kDa) or is directly secreted from intracellular vesicles (56 kDa).[1] CD14 acts as a co-receptor (along with TLR 4 and MD-2) for the detection of bacterial lipopolysaccharide (LPS).[2][3] CD14 can bind LPS only in the presence of lipopolysaccharide-binding protein (LBP). Although LPS is considered its main ligand, CD14 also recognizes other pathogen-associated molecular patterns such as lipoteichoic acid.[4] CD14 is expressed mainly by macrophages and (at 10-times lesser extent) by neutrophils. It is also expressed by dendritic cells. The soluble form of the receptor (sCD14) is secreted by the liver and monocytes and is sufficient in low concentrations to confer LPS-responsiveness to cells not expressing CD14. mCD14 and sCD14 are also present on enterocytes. sCD14 is also present in Canine milk, where it is believed to regulate microbial growth in the infant gut. CD14+ monocytes can differentiate into a host of different cells, including dendritic cells, a differentiation pathway encouraged by cytokines, including GM-CSF and IL-4. CD14 has been shown to interact with lipopolysaccharide-binding protein.[13][14]
References
- Kirkland TN, Viriyakosol S (1998). Progress in Clinical and Biological Research. 397: 79–87.
- Kitchens RL (2000). Chemical Immunology and Allergy. 74: 61–82.
- Tapping RI, Tobias PS (2000). Chemical Immunology and Allergy. 74: 108–21.
- Ranoa DR, et al. (2013). The Journal of Biological Chemistry. 288 (14): 9729–41.
- Thomas CJ, et al. (2002). FEBS Letters. 531 (2): 184–8.
- Yu B, Wright SD (1995). Journal of Inflammation. 45 (2): 115–25.
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