Human alpha 1 Antitrypsin ELISA Kit, Fluorescent (ab229417)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 5.8 ng/ml
- Range: 7.8 ng/ml - 2000 ng/ml
- Sample type: Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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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.
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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.
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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 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 Antibody Diluent 4BI 1 x 6ml Human alpha 1 Antitrypsin Lyophilized Purified Protein 2 vials 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
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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
modificationsN-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
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Alternative names
- A1A
- A1AT
- A1AT_HUMAN
see all -
Database links
- Entrez Gene: 5265 Human
- Omim: 107400 Human
- SwissProt: P01009 Human
- Unigene: 525557 Human
Images
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Other - Human alpha 1 Antitrypsin ELISA Kit, Fluorescent (ab229417)
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 alpha 1 Antitrypsin (SERPINA1) standard curve in Sample Diluent NSExample 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.
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Titration of Human serum within the working range of the assayBackground-subtracted data values (mean +/- SD, n = 2) are graphed.
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Observed alpha 1 Antitrypsin levels in individual donor normal Human serum (n=10)The mean alpha 1 Antitrypsin concentration was determined to be 72 mg/mL with a range of 17-181 mg/mL.
