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
製品の概要
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製品名
Human alpha 1 Antitrypsin ELISA Kit (SERPINA1)
alpha 1 Antitrypsin キット 製品一覧 -
検出方法
Colorimetric -
再現性
Intra-Assay(同時再現性) サンプル N 平均値 SD CV% Overall 5 3.9% Inter-Assay(日差再現性) サンプル N 平均値 SD CV% Overall 3 2.1% -
サンプルの種類
Serum, Hep Plasma, EDTA Plasma, Cit plasma -
アッセイタイプ
Sandwich (quantitative) -
検出感度
13 ng/ml -
検出範囲
31.25 ng/ml - 2000 ng/ml -
添加回収試験
特定サンプルでの回収試験 サンプルの種類 平均 % 測定範囲 Serum 96 92% - 101% Hep Plasma 99 94% - 102% EDTA Plasma 77 70% - 87% Cit plasma 85 83% - 86% -
全工程の試験時間
1h 30m -
ステップ
One step assay -
種交差性
交差種: Human
非交差種: Goat, Cow, Pig, Cynomolgus monkey, Macaque monkey -
製品の概要
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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特記事項
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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試験プラットフォーム
Microplate
製品の特性
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保存方法
Store at +4°C. Please refer to protocols. -
内容 1 x 96 tests 10 x 96 tests 10X Human alpha 1 Antitrypsin Capture Antibody 1 x 600µl 10 x 600µl 10X Human alpha 1 Antitrypsin Detector Antibody 1 x 600µl 10 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 1 x 200ml Antibody Diluent 4BI 1 x 6ml 10 x 6ml Human alpha 1 Antitrypsin Lyophilized Purified Protein 2 vials 20 vials Plate Seals 1 unit 10 units Sample Diluent NS (ab193972) 1 x 50ml 2 x 250ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit 10 units Stop Solution 1 x 12ml 1 x 120ml TMB Development Solution 1 x 12ml 1 x 120ml -
研究分野
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機能
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). -
組織特異性
Ubiquitous. Expressed in leukocytes and plasma. -
関連疾患
Alpha-1-antitrypsin deficiency -
配列類似性
Belongs to the serpin family. -
ドメイン
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. -
翻訳後修飾
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. -
細胞内局在
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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別名
- A1A
- A1AT
- A1AT_HUMAN
see all -
参照データベース
- Entrez Gene: 5265 Human
- Omim: 107400 Human
- SwissProt: P01009 Human
- Unigene: 525557 Human
関連製品
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ELISA kits
画像
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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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Background-subtracted data values (mean +/- SD) are graphed.
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Example 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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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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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 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.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (7)
ab189579 は 7 報の論文で使用されています。
- 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