Overview

  • Product name

    Human AACT ELISA Kit
    See all AACT kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Plasma 5 4.3%
    Inter-assay
    Sample n Mean SD CV%
    Plasma 3 4%
  • Sample type

    Cell culture supernatant, Milk, Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    327 pg/ml
  • Range

    1176 pg/ml - 40000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 108 96% - 118%
    Cell culture extracts 98 91% - 105%
    Tissue Extracts 96 87% - 102%
    Cell culture media 109 103% - 112%
    Heparin Plasma 108 101% - 115%
    EDTA Plasma 106 102% - 111%
    Citrate Plasma 99 84% - 112%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    Human AACT SimpleStep ELISA® kit (ab171574) has been re-developed with new capture and detector antibodies. This new kit has the same name but a different product number (ab217779). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantifying AACT in human serum, citrate plasma, EDTA plasma, heparin plasma, cell culture media and extracts and tissue extracts.


    AACT in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of AACT protein in serum, plasma, milk, cell culture supernatant, cell and tissue extract. 


    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 substrate 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.


    Sensitivity:


    Samples in 1X Cell Extraction Buffer PTR: 327 pg/mL


    Samples in Sample Diluent NS: 608 pg/mL

  • Notes

    AACT (Alpha 1-antichymotrypsin) can inhibit neutrophil cathepsin G and mast cell chymase, both of which can convert angiotensin-1 to the active angiotensin-2.  AACT is synthesized in the liver and like the related alpha-1-antitrypsin, its concentration increases in the acute phase of inflammation or infection.  AACT is also found in the amyloid plaques from the hippocampus of Alzheimer disease brains.  Defects in AACT may be a cause of chronic obstructive pulmonary disease (COPD).

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human AACT Capture Antibody 1 x 600µl
    10X Human AACT 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 CPI 1 x 6ml
    Human AACT Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Although its physiological function is unclear, it can inhibit neutrophil cathepsin G and mast cell chymase, both of which can convert angiotensin-1 to the active angiotensin-2.
  • Tissue specificity

    Plasma. Synthesized in the liver. Like the related alpha-1-antitrypsin, its concentration increases in the acute phase of inflammation or infection. Found in the amyloid plaques from the hippocampus of Alzheimer disease brains.
  • Involvement in disease

    Defects in SERPINA3 may be a cause of chronic obstructive pulmonary disease (COPD) [MIM:107280].
  • Sequence similarities

    Belongs to the serpin family.
  • Domain

    The reactive center loop (RCL) extends out from the body of the protein and directs binding to the target protease. The protease cleaves the serpin at the reactive site within the RCL, establishing a covalent linkage between the carboxyl group of the serpin reactive site and the serine hydroxyl of the protease. The resulting inactive serpin-protease complex is highly stable.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • SERPINA3
    • AACT
    • AACT_HUMAN
    • ACT
    • alpha 1 Antichymotrypsin
    • Alpha-1-antichymotrypsin His-Pro-less
    • Antichymotrypsin
    • Cell growth inhibiting gene 24/25 protein
    • Cell growth-inhibiting gene 24/25 protein
    • GIG24
    • GIG25
    • Growth inhibiting protein 24
    • Growth inhibiting protein 25
    • MGC88254
    • Serine (or cysteine) proteinase inhibitor clade A (alpha 1 antiproteinase, antitrypsin) member 3
    • Serine (or cysteine) proteinase inhibitor clade A member 3
    • Serine proteinase inhibitor clade A member 3
    • Serpin A3
    • Serpin family A member 3
    • Serpin peptidase inhibitor clade A (alpha 1 antiproteinase antitrypsin) member 3
    • SERPINA3
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab217779 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.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Alpha 1-antichymotrypsin were measured in duplicates, interpolated from the Alpha 1-antichymotrypsin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:10,000, plasma (citrate) 1:10,000, plasma (heparin) 1:10,000 and plasma (EDTA) 1:10,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Alpha 1-antichymotrypsin concentration was determined to be 310 µg/mL in serum, 323 µg/mL in plasma (citrate) 248 µg/mL in plasma (heparin) and 254 µg/mL in plasma (EDTA).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Alpha 1-antichymotrypsin concentration was determined to be 266 µg/mL with a range of 190 – 364 µg/mL.

  • The concentrations of Alpha 1-antichymotrypsin were measured in duplicates, interpolated from the Alpha 1-antichymotrypsin standard curves and corrected for sample dilution. Undiluted samples are as follows: milk 1:500 and HEPG2 supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Alpha 1-antichymotrypsin concentration was determined to be 14,672 ng/mL in milk and 60 ng/mL in HEPG2 cell culture supernatant.

  • The concentrations of Alpha 1-antichymotrypsin were measured in duplicate and interpolated from the Alpha 1-antichymotrypsin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Alpha 1-antichymotrypsin concentration was determined to be 144 ng/mL in HEPG2 cell extract and 56 ng/mL in human liver homogenate extract.

Protocols

References

ab217779 has not yet been referenced specifically in any publications.

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