Human LCAT ELISA Kit is a single-wash 90-min Simplestep used to quantify Human LCAT with a sensitivity of 6.22 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
Images
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
- 31.25 - 2000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 6.22 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Target data
Function
Central enzyme in the extracellular metabolism of plasma lipoproteins. Synthesized mainly in the liver and secreted into plasma where it converts cholesterol and phosphatidylcholines (lecithins) to cholesteryl esters and lysophosphatidylcholines on the surface of high and low density lipoproteins (HDLs and LDLs) (PubMed:10329423, PubMed:19065001, PubMed:26195816). The cholesterol ester is then transported back to the liver. Has a preference for plasma 16:0-18:2 or 18:O-18:2 phosphatidylcholines (PubMed:8820107). Also produced in the brain by primary astrocytes, and esterifies free cholesterol on nascent APOE-containing lipoproteins secreted from glia and influences cerebral spinal fluid (CSF) APOE- and APOA1 levels. Together with APOE and the cholesterol transporter ABCA1, plays a key role in the maturation of glial-derived, nascent lipoproteins. Required for remodeling high-density lipoprotein particles into their spherical forms (PubMed:10722751). Catalyzes the hydrolysis of 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine (platelet-activating factor or PAF) to 1-O-alkyl-sn-glycero-3-phosphocholine (lyso-PAF) (PubMed:8016111). Also catalyzes the transfer of the acetate group from PAF to 1-hexadecanoyl-sn-glycero-3-phosphocholine forming lyso-PAF (PubMed:8016111). Catalyzes the esterification of (24S)-hydroxycholesterol (24(S)OH-C), also known as cerebrosterol to produce 24(S)OH-C monoesters (PubMed:24620755).
Alternative names
Phosphatidylcholine-sterol acyltransferase, 1-alkyl-2-acetylglycerophosphocholine esterase, Lecithin-cholesterol acyltransferase, Phospholipid-cholesterol acyltransferase, Platelet-activating factor acetylhydrolase, PAF acetylhydrolase, LCAT
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Human LCAT ELISA Kit is a single-wash 90-min Simplestep used to quantify Human LCAT with a sensitivity of 6.22 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
- 31.25 - 2000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 6.22 pg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Serum | n 8 | mean - | SD - | C.V. 6.2 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Serum | n 3 | mean - | SD - | C.V. 3.1 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture supernatant | Average % = 99 | Range 95 - 104 % |
Sample type Serum | Average % = 80 | Range 72 - 84 % |
Sample type EDTA Plasma | Average % = 85 | Range 78 - 91 % |
Sample type Heparin Plasma | Average % = 105 | Range 103 - 108 % |
Sample type Citrate plasma | Average % = 120 | Range 115 - 128 % |
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 LCAT ELISA Kit (ab288181) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Human LCAT protein in plasma, cell culture supernatant and serum samples. It uses our proprietary SimpleStep ELISA® technology. Quantitate LCAT with 6.22 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
LCAT also known as lecithin-cholesterol acyltransferase is an enzyme with a molecular mass of approximately 68 kDa. It belongs to the family of esterases and is mainly expressed in the liver. LCAT circulates in the bloodstream playing a significant role in lipid metabolism by catalyzing the formation of cholesteryl esters from free cholesterol and phosphatidylcholine (lecithin) on the surface of high-density lipoproteins (HDL).
- Biological function summary
This enzyme functions centrally in the maturation of high-density lipoprotein particles. LCAT interacts with and acts upon HDL converting nascent HDL into mature HDL by catalyzing the esterification of cholesterol. This process results in the sequestration of cholesterol within the core of HDL particles essential for reverse cholesterol transport to the liver. Moreover LCAT does not operate as part of a larger complex but is an active enzyme on its own within various lipid transport processes.
- Pathways
LCAT's action significantly influences the lipid metabolism and reverse cholesterol transport pathways. In the lipid metabolism pathway it works closely with apolipoproteins such as apoA-I which serves as an activator. ApoA-I stabilizes the enzymatic action of LCAT promoting cholesterol homeostasis whereas LCAT ensures the efficient transfer of cholesterol to the liver within the reverse cholesterol transport pathway contributing to the maintenance of lipid balance.
- Associated diseases and disorders
LCAT deficiency can lead to conditions like familial LCAT deficiency and Fish Eye disease. These disorders are marked by abnormal cholesterol and lipid accumulation due to impaired LCAT activity. Apolipoproteins particularly apoA-I also demonstrate a significant connection as mutations or lack of apoA-I compromise LCAT activation exacerbating hyperlipidemia and related lipid disorders.
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4 product images
Sandwich ELISA - Human LCAT ELISA Kit (ab288181)
Interpolated concentrations of LCAT in normal human serum donors. Serum of ten individual healthy human female donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +- SD, n=2). The mean LCAT concentration was determined to be 4.5 ug/mL with a range of 3.7 - 6.5 ug/mL.
Sandwich ELISA - Human LCAT ELISA Kit (ab288181)
Interpolated concentrations of human LCAT in HepG2 cell culture supernatant. Interpolated concentration of native LCAT was measured in duplicate at different sample concentrations. Undiluted samples are as follows: HepG2 cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Sandwich ELISA - Human LCAT ELISA Kit (ab288181)
Interpolated concentrations of human LCAT in serum, plasma (heparin), plasma (EDTA), and plasma (citrate). Interpolated concentration of native LCAT was measured in duplicate at different sample concentrations. Undiluted samples are as follows: serum 1:4,000, plasma (heparin) 1:2,000, plasma (EDTA) 1:2,000, and plasma (citrate) 1:5,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS
Sandwich ELISA - Human LCAT ELISA Kit (ab288181)
Example of human LCAT standard curve in Sample Diluent NS. Example of human LCAT standard curve. Background-subtracted data values (mean +/- SD) are graphed.
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