Overview

  • Product name

    Human alpha 1 Antitrypsin ELISA Kit, Fluorescent
    See all alpha 1 Antitrypsin kits
  • Detection method

    Fluorescent
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Human serum 5 3.9%
    Inter-assay
    Sample n Mean SD CV%
    Human Serum 3 2.1%
  • Sample type

    Serum, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    5.8 ng/ml
  • Range

    7.8 ng/ml - 2000 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 96 92% - 101%
    Hep Plasma 99 94% - 102%
    EDTA Plasma 77 70% - 87%
    Cit plasma 85 83% - 86%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Goat, Cow, Pig
  • Product overview

    alpha 1 Antitrypsin (SERPINA1) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of alpha 1 Antitrypsin (SERPINA1) protein in human serum and plasma.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint 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. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    Alpha 1 Antitrypsin (SERPINA1) is a serine protease inhibitor found in a variety of biological fluids. While it can inhibit trypsin, chymotrypsin, and plasminogen activator, its main target is elastase. Alpha 1 Antitrypsin plays a critical role in the lung by inhibiting the activity of elastase, produced by neutrophils during inflammatory responses. The aberrant form plays a role in coagulation and insulin induced NO synthesis. Alpha 1 Antitrypsin deficiency results in emphysema and may also result in liver disease.

  • 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
    100X Stoplight Red Substrate 1 x 120µl
    10X Human alpha 1 Antitrypsin Capture Antibody 1 x 600µl
    10X Human alpha 1 Antitrypsin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Human alpha 1 Antitrypsin Lyophilized Purified Protein 2 vials
    Antibody Diluent 4BI 1 x 6ml
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas

  • Function

    Inhibitor of serine proteases. Its primary target is elastase, but it also has a moderate affinity for plasmin and thrombin. Irreversibly inhibits trypsin, chymotrypsin and plasminogen activator. The aberrant form inhibits insulin-induced NO synthesis in platelets, decreases coagulation time and has proteolytic activity against insulin and plasmin.
    Short peptide from AAT: reversible chymotrypsin inhibitor. It also inhibits elastase, but not trypsin. Its major physiological function is the protection of the lower respiratory tract against proteolytic destruction by human leukocyte elastase (HLE).
  • Tissue specificity

    Ubiquitous. Expressed in leukocytes and plasma.
  • Involvement in disease

    Alpha-1-antitrypsin deficiency
  • 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.
  • Post-translational
    modifications

    N-glycosylated. Differential glycosylation produces a number of isoforms. N-linked glycan at Asn-107 is alternatively di-antennary, tri-antennary or tetra-antennary. The glycan at Asn-70 is di-antennary with trace amounts of tri-antennary. Glycan at Asn-271 is exclusively di-antennary. Structure of glycans at Asn-70 and Asn-271 is Hex5HexNAc4. The structure of the antennae is Neu5Ac(alpha1-6)Gal(beta1-4)GlcNAc attached to the core structure Man(alpha1-6)[Man(alpha1-3)]Man(beta1-4)GlcNAc(beta1-4)GlcNAc. Some antennae are fucosylated, which forms a Lewis-X determinant.
    Proteolytic processing may yield the truncated form that ranges from Asp-30 to Lys-418.
  • Cellular localization

    Secreted. Endoplasmic reticulum. The S and Z allele are not secreted effectively and accumulate intracellularly in the endoplasmic reticulum and Secreted, extracellular space, extracellular matrix.
  • Information by UniProt
  • Alternative names

    • A1A
    • A1AT
    • A1AT_HUMAN
    • AAT
    • Alpha 1 antiproteinase
    • Alpha 1 antitrypsin
    • Alpha 1 antitrypsin null
    • Alpha 1 protease inhibitor
    • Alpha-1 protease inhibitor
    • Alpha-1-antiproteinase
    • alpha1 proteinase inhibitor
    • Alpha1AT
    • Dom1
    • PI
    • PI1
    • PRO0684
    • PRO2275
    • Serine (or cysteine) proteinase inhibitor clade A member 1
    • Serine protease inhibitor 1-1
    • Serine protease inhibitor A1a
    • Serpin A1
    • Serpin A1a
    • Serpin peptidase inhibitor clade A member 1
    • Serpina1
    • Short peptide from AAT
    • SPAAT
    • Spi1-1
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229417 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.

     

  • Example of human alpha 1 Antitrypsin (SERPINA1) standard curve in Sample Diluent NS. The alpha 1 Antitrypsin (SERPINA1) standard curve was prepared as described in Section 10.

  • Background-subtracted data values (mean +/- SD, n = 2) are graphed.

  • The mean alpha 1 Antitrypsin concentration was determined to be 72 mg/mL with a range of 17-181 mg/mL.

Protocols

References

ab229417 has not yet been referenced specifically in any publications.

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