Human LC3A ELISA Kit (ab239432)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 2.1 pg/ml
- Range: 39.06 pg/ml - 2500 pg/ml
- Sample type: Cell culture extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human LC3A ELISA Kit
See all LC3A kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% cell extract 8 3.6% Inter-assay Sample n Mean SD CV% cell extract 3 3.5% -
Sample type
Cell culture extracts -
Assay type
Sandwich (quantitative) -
Sensitivity
2.1 pg/ml -
Range
39.06 pg/ml - 2500 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture extracts 103 97% - 109% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human LC3A ELISA Kit (ab239432) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of LC3A protein in cell culture extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human LC3A with 2.1 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 stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human LC3A Capture Antibody 1 x 600µl 10X Human LC3A Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BI 1 x 6ml Human LC3A Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Relevance
A major contributor to cellular homeostasis is the ability of the cell to strike a balance between the formation and degradation/removal of its cellular components. This process of internal cellular turn-over is called autophagy (self-eating), and is facilitated by a pathway of around 16 interacting proteins in the human. LC3, a ubiquitin-like modifier protein, is the human homolog of yeast Apg8 and is involved in the formation of autophagosomal vacuoles, called autophagosomes. LC3 is expressed as 3 splice variants (LC3A, LC3B and LC3C), which exhibit different tissue distributions and are processed into cytosolic and autophagosomal membrane-bound forms, termed LC3-I and LC3-II, respectively. A disruption to the autophagic process is now associated with the progression of several cancers, neurodegenerative disorders and cardiac pathologies, where LC3 is widely employed as a marker for autophagy. -
Cellular localization
LC3-I is located in the Cytoplasm. LC3-II is found on intracytoplasmic membranes and the autophagosomal membrane. -
Alternative names
- ATG 8
- Autophagy related ubiquitin-like modifier ATG8
- Autophagy-related protein 8
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Database links
- Entrez Gene: 440738 Human
- Omim: 609605 Human
- SwissProt: Q9BXW4 Human
Images
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Other - Human LC3A ELISA Kit (ab239432)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human LC3A standard curve in 1X Cell Extraction Buffer PTR.The LC3A standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed. -
Interpolated concentrations of native LC3A in human SHSY-5Y cell extract based on a 30 µg/mL extract load, and HeLa cell extract based on a 500 µg/mL extract load.The concentrations of LC3A were measured in duplicate and interpolated from the LC3A standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean LC3A concentration was determined to be 2,048 pg/mL in SHSY-5Y cell extract and 1,601 pg/mL in HeLa cell extract. -
Comparison of LC3A in mock-treated and chloroquine-treated HeLa cell extracts.HeLa cells were cultured for 24 hours in the absence (mock) or presence of 50 mM chloroquine. The concentrations of LC3A were measured in three different dilutions of the supernatant samples in duplicates and interpolated from the LC3A standard curve. The interpolated values are plotted (mean +/- SD, n=3). The mean LC3A concentration was determined to be 3.16 ng/mg in mock-treated HeLa cell extract, and 5.64 ng/mg in chloroquine-treated HeLa cell extract. -
Sandwich ELISA - Human LC3A ELISA Kit (ab239432)To learn more about the advantages of recombinant antibodies see here.
