Human alpha 1 Antitrypsin ELISA Kit (SERPINA1) (ab189579)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 13 ng/ml
- Range: 31.25 ng/ml - 2000 ng/ml
- Sample type: Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human alpha 1 Antitrypsin ELISA Kit (SERPINA1)
See all alpha 1 Antitrypsin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Overall 5 3.9% Inter-assay Sample n Mean SD CV% Overall 3 2.1% -
Sample type
Serum, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
13 ng/ml -
Range
31.25 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, Cynomolgus monkey, Macaque monkey -
Product overview
Human alpha 1 Antitrypsin ELISA Kit (SERPINA1) (ab189579) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of alpha 1 Antitrypsin (SERPINA1) protein in cit plasma, hep plasma, serum, and edta plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human alpha 1 Antitrypsin (SERPINA1) with 13 ng/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 strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
ASSAY SPECIFICITY
This kit recognizes both native and recombinant human alpha 1 Antitrypsin protein in serum, plasma, and cell culture supernatant, cell and tissue extract samples only.
Normal human urine and saliva samples tested negative for this kit.
CROSS REACTIVITY
Human alpha 1 Anti-chymotrypsin protein and human PAI protein were prepared at 2,000 ng/mL and 1,000 ng/mL respectively and assayed for cross reactivity. No cross-reactivity was observed.
INTERFERENCE
Human alpha 1 Anti-chymotrypsin protein and human PAI protein were prepared at 2,000 ng/mL and 1,000 ng/mL respectively and assayed for and tested for interference. No interference with was observed.
SPECIES REACTIVITY
This kit recognizes human alpha 1 Antitrypsin protein.
Other species reactivity was determined by measuring alpha 1 Antitrypsin (1:10,000) serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution.
Reactivity < 3% was determined for the following species: Mouse, Rat, Hamster, Guinea Pig, Rabbit, Dog, Goat, Pig and Cow.
CALIBRATION
This immunoassay is calibrated against a highly purified human alpha 1 Antitrypsin. The NIBSC/WHO unclassified purified human alpha 1 Antitrypsin preparation 05/162 was evaluated in this kit.
The dose response curve of the unclassified standard alpha 1 Antitrypsin parallels the SimpleStep standard curve. To convert sample values obtained with the SimpleStep Human alpha 1 Antitrypsin kit to approximate NIBSC 05/162 units, use the equation below.
NIBSC (05/162) approximate value (ng/mL) = 0.3649 x SimpleStep Human alpha 1 Antitrypsin value (ng/mL).
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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, it’s 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
Microplate
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 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 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 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 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
Associated products
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ELISA kits
Images
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Other - Human alpha 1 Antitrypsin ELISA Kit (SERPINA1) (ab189579)
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 alpha 1 Antitrypsin standard curve.Background-subtracted data values (mean +/- SD) are graphed.
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Standard curveExample of human alpha 1 Antitrypsin standard curve in Sample Diluent NS. The alpha 1 Antitrypsin standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native alpha 1 Antitrypsin in human serum and plasma samples.The concentrations of alpha 1 Antitrypsin were measured in duplicates, interpolated from the alpha 1 Antitrypsin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 0.005%, plasma (citrate) 0.025%, plasma (heparin) 0.005% ,and plasma (EDTA) 0.005%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean alpha 1 Antitrypsin concentration was determined to be 30 mg/mL in serum, 5 mg/mL in plasma (citrate), 19 mg/mL in plasma (heparin) and 21 mg/mL in in plasma (EDTA).
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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.
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Linearity of dilution.Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.
Native alpha 1 Antitrypsin was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab189579 has been referenced in 7 publications.
- Rotondo JC et al. SERPINA1 Gene Promoter Is Differentially Methylated in Peripheral Blood Mononuclear Cells of Pregnant Women. Front Cell Dev Biol 8:550543 (2020). PubMed: 33015055
- Du S et al. Serum alpha 1-antitrypsin predicts severe acute kidney injury after cardiac surgery. J Thorac Dis 11:5053-5062 (2019). PubMed: 32030221
- Lin H et al. a1-Anti-trypsin improves function of porcine donor lungs during ex-vivo lung perfusion. J Heart Lung Transplant 37:656-666 (2018). PubMed: 29153638
- Hansen HG et al. Case study on human a1-antitrypsin: Recombinant protein titers obtained by commercial ELISA kits are inaccurate. Biotechnol J 11:1648-1656 (2016). Sandwich ELISA ; Human . PubMed: 27731958
- Hennawy MG et al. The effect of a1-antitrypsin deficiency combined with increased bacterial loads on chronic obstructive pulmonary disease pharmacotherapy: A prospective, parallel, controlled pilot study. J Adv Res 7:1019-1028 (2016). Sandwich ELISA ; Human . PubMed: 27857848
- Kim S & Lee HJ Direct Detection of alpha-1 Antitrypsin in Serum Samples using Surface Plasmon Resonance with a New Aptamer-Antibody Sandwich Assay. Anal Chem N/A:N/A (2015). ELISA ; Human . PubMed: 26070325
- Laurent SA et al. ?-secretase directly sheds the survival receptor BCMA from plasma cells. Nat Commun 6:7333 (2015). ELISA ; Human . PubMed: 26065893