Rat Alpha-1-Antitrypsin (a1AT) ELISA Kit

Due to the possibility of mismatching between antigens from other origin and antibodies used in our kits (e.g., antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.

Principle of the Assay

The microtiter plate provided in this kit has been pre-coated with an antibody specific to a1AT. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to a1AT. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain a1AT, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution, and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of a1AT in the samples is then determined by comparing the O.D. of the samples to the standard curve.

For Use with serum, plasma, and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

Inhibitor of serine proteases. The primary target is elastase, but also has a moderate affinity for plasmin and thrombin.

GENE ID	24648
SWISS PROT	P17475
SYNONYMS	SERPINA1; SPAAT; A1-AT; PI; Serpin Peptidase Inhibitor,Clade A(Alpha-1 Antiproteinase/AntiTrypsin)Member 1; Alpha-1 protease inhibitor; Short peptide from AAT; Serpin A1

Materials Supplied

Kit Components	96 Wells Quantity/Size
Pre-coated, ready-to-use 96-well strip plate	1 plate
Plate sealer for 96 wells	2
Standard	2 tubes
Diluent buffer	1 bottle
Detection Reagent A	1 bottle
Detection Reagent B	1 bottle
TMB Substrate	1 tube
Stop Solution	1 tube
Wash Buffer (30 ℅ concentrate)	1 tube
Product data sheet	1 copy

Storage

The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C.

Performance Characteristics

REPEATABILITY	Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level a1AT were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle, and high-level a1AT were tested on 3 different plates, with 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
SENSITIVITY	The minimum detectable dose was 29ng/mL.
ASSAY RANGE	78.1-5000ng/mL
SPECIFICITY	This assay has high sensitivity and excellent specificity for the detection of a1AT. No significant cross-reactivity or interference between a1AT and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between a1AT and all analogs, therefore, cross reactivity may still exist.