Overview

  • 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

    LC3A in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of LC3A protein in human cell and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    LC3A is a ubiquitin-like modifier involved in formation of autophagosomal vacuoles (autophagosomes). LC3s are involved in elongation of the phagophore membrane. The precursor molecule is cleaved by ATG4B to form the cytosolic form, LC3-I. This form is activated by APG7L/ATG7, transferred to ATG3 and conjugated to phospholipid to form the membrane-bound form, LC3-II.


     

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab239432 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

  • 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.
  • 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.
  • 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.

Protocols

References

ab239432 has not yet been referenced specifically in any publications.

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