Human CD14 ELISA Kit is a single-wash 90-min Simplestep used to quantify Human CD14 with a sensitivity of 4 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
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Key facts
- Detection method
- Colorimetric
- Sample types
- Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 31.3 - 2000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 4 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Coreceptor for bacterial lipopolysaccharide (PubMed:1698311, PubMed:23264655). In concert with LBP, binds to monomeric lipopolysaccharide and delivers it to the LY96/TLR4 complex, thereby mediating the innate immune response to bacterial lipopolysaccharide (LPS) (PubMed:20133493, PubMed:22265692, PubMed:23264655). Acts via MyD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response (PubMed:8612135). Acts as a coreceptor for TLR2:TLR6 heterodimer in response to diacylated lipopeptides and for TLR2:TLR1 heterodimer in response to triacylated lipopeptides, these clusters trigger signaling from the cell surface and subsequently are targeted to the Golgi in a lipid-raft dependent pathway (PubMed:16880211). Binds electronegative LDL (LDL(-)) and mediates the cytokine release induced by LDL(-) (PubMed:23880187).
Alternative names
CD14, Monocyte differentiation antigen CD14, My23 antigen, Myeloid cell-specific leucine-rich glycoprotein
What's included?
Recommended products
Human CD14 ELISA Kit is a single-wash 90-min Simplestep used to quantify Human CD14 with a sensitivity of 4 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
Key facts
- Detection method
- Colorimetric
- Sample types
- Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 31.3 - 2000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Microplate (12 x 8 well strips)
- Sensitivity
- = 4 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Human Serum | n 8 | C.V. 2.8 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Human Serum | n 3 | C.V. 2.1 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture supernatant | Average % = 102 | Range 95 - 106 % |
Sample type Serum | Average % = 110 | Range 104 - 114 % |
Sample type EDTA Plasma | Average % = 110 | Range 105 - 115 % |
Sample type Heparin Plasma | Average % = 109 | Range 106 - 111 % |
Sample type Citrate plasma | Average % = 109 | Range 99 - 116 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Human CD14 ELISA Kit (ab208983) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of CD14 protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human CD14 with 4 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
CD14 is a LPS-binding glycoprotein primarily expressed on myeloid cells. In response to a gram negative bacterial infection, LPS-activated CD14 leads to the production of inflammatory cytokines. CD14 plays a role in additional cellular functions, including recognition of peptidylglycan from gram positive cells and binding of apoptotic cells to induce phagocytosis.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
CD14 also known as cluster of differentiation 14 is a glycoprotein with a molecular mass of approximately 55 kDa. This protein is expressed mainly on the surface of macrophages monocytes and to a lesser extent on neutrophils. CD14 exists in two forms: membrane-bound (mCD14) and soluble (sCD14). It functions as a co-receptor along with TLR4 (Toll-like receptor 4) and MD-2 for the detection of bacterial lipopolysaccharide (LPS) which is an important component of the outer membrane of Gram-negative bacteria.
- Biological function summary
This glycoprotein plays a significant role in the innate immune system by recognizing pathogen-associated molecular patterns (PAMPs). It is part of a receptor complex where CD14 acts alongside LPS binding protein (LPS-R) TLR4 and MD-2 to facilitate pro-inflammatory signaling in response to microbial invasion. By binding to LPS CD14 initiates and amplifies the immune response leading to the production of cytokines and the recruitment of other immune cells to the site of infection.
- Pathways
CD14 interacts with the toll-like receptor signaling pathway notably involving TLR4. Upon LPS detection CD14 assists in the activation of TLR4 triggering downstream signaling cascades such as NF-κB and MAPK pathways. These pathways result in the transcription of inflammatory cytokines and are essential for the host defense mechanism. The protein complex involving CD14 further interacts with adaptor proteins like MyD88 and TRIF facilitating signal propagation and the immune system's ability to respond to pathogens.
- Associated diseases and disorders
Researchers have linked CD14 to sepsis and atherosclerosis. High levels of soluble CD14 (sCD14) are often observed in patients with sepsis indicating its participation in excessive systemic inflammation. In atherosclerosis CD14's role in recognizing LPS contributes to chronic inflammation and the development of plaques in arteries. Through these diseases CD14 connects with TLR4 emphasizing its impact on inflammatory responses and its potential as a therapeutic target.
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5 product images
Sandwich ELISA - Human CD14 ELISA Kit (ab208983)
House CD14 standard curve comparison data.
Standard curve comparison between human CD14 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 30-fold increase in sensitivity.
Sandwich ELISA - Human CD14 ELISA Kit (ab208983)
Example of human CD14 standard curve in Sample Diluent NS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human CD14 ELISA Kit (ab208983)
Interpolated concentrations of native CD14 in human serum and plasma samples.
The concentrations of CD14 were measured in duplicates, interpolated from the CD14 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:625, plasma (citrate) 1:800, plasma (heparin) 1:800 and plasma (EDTA) 1:800. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD14 concentration was determined to be 897 ng/mL in serum, 1128 ng/mL in plasma (citrate), 1,195 ng/mL in plasma (heparin) and 1,212 ng/mL in plasma (EDTA).
Sandwich ELISA - Human CD14 ELISA Kit (ab208983)
Interpolated concentrations of native CD14 in human stimulated and unstimulated PBMC.
The concentrations of CD14 were measured, interpolated from the CD14 standard curves and corrected for sample dilution. Undiluted samples are as follows: PHA stimulated PBMC 25% and unstimulated PBMC 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD14 concentration was determined to be 7.91 ng/mL in PHA stimulated PBMC and 6.92 ng/mL in unstimulated PBMC.
Sandwich ELISA - Human CD14 ELISA Kit (ab208983)
Serum from ten individual healthy human male donors was measured in duplicate.
Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD14 concentration was determined to be 1,360 ng/mL with a range of 429 – 2,955 ng/mL.
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