LKM-1 Ab ELISA Kit

Sample

Serum

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The LKM-1 Ab ELISA Kit is a solid phase enzyme immunoassay employing human recombinant cytochrome p450 IID6 for the quantitative and qualitative detection of antibodies against liver-kidney microsomes (LKM) in human serum.

Contents of Kit

1. Sample Buffer (5×): 5× concentrated Tris, sodium chloride (NaCl), bovine serum albumin (BSA), sodium azide < 0.1% (preservative). Yellow, 20 mL
2. Wash Buffer (50×): 50× concentrated Tris, NaCl, Tween 20, sodium azide < 0.1% (preservative). Green, 20 mL
3. Negative Control (Control 2): Ready to use. Human serum (diluted), bovine serum albumin (BSA), sodium azide < 0.1% (preservative). Colorless, 1.5 mL
4. Positive Control (Control 1): Ready to use. Human serum (diluted), bovine serum albumin (BSA), sodium azide < 0.1% (preservative). Yellow, 1.5 mL
5. Cut-off Control (Control 3): Ready to use. Human serum (diluted), bovine serum albumin (BSA), sodium azide < 0.1% (preservative). Yellow 1.5 mL
6. Calibrator A-F: Ready to use. Concentration of each calibrator: 0, 3, 10, 30, 100, 300 U/mL. Human serum (diluted), bovine serum albumin (BSA), sodium azide < 0.1% (preservative). Yellow (Color increasing with concentration), 1.5 mL × 6
7. Conjugate IgG: Ready to use. Anti-human immunoglobulins conjugated to horseradish peroxidase, bovine serum albumin (BSA). Blue, 15 mL
8. TMB Substrate: Ready to use. Stabilized tetramethylbenzidine and hydrogen peroxide (TMB/H₂O₂). Colorless, 15 mL
9. Stop Solution: Ready to use. 1 M Hydrochloric Acid. Colorless, 15 mL
10. Microtiter Plate Ready to use. With breakaway microwells. 12 × 8 well strips

Storage

Store all reagents and the microplate at 2-8°C, in their original containers. Once prepared, reconstituted solutions are stable at 2-8°C for 1 month, at least. Reagents and the microplate shall be used within the expiry date indicated on each component, only. Avoid intense exposure of TMB solution to light. Store microplates in designated foil, including the desiccant, and seal tightly.

Performance Characteristics

Calibration range: 0-300 U/mL
Due the lack of international reference calibration this assay is calibrated in arbitrary units (U/mL).

Precision

To determine the precision of the assay, the variability (intra and inter-assay) was assessed by examining its reproducibility on three serum samples selected to represent a range over the standard curve.

Sensitivity

Testing sample buffer 30 times on LKM-1 Ab ELISA Kit gave a sensitivity of 1.0 U/mL.

General Description

Autoimmune hepatitis (AIH) is a chronic progressive liver disease of unknown origin that responds well to immunosuppressive therapy, but has a poor prognosis if untreated. Early and accurate diagnosis is therefore of great importance. AIH is characterized by histological features of periportal hepatitis in the absense of viral markers, by hypergammaglobulinemia and, in the majority of patients, by the presence of autoantibodies in serum. Anti-nuclear antibodies (ANA), smooth muscle antibodies (SMA), anti-liver kidney microsomal antibodies (LKM) and antibodies against soluble liver antigen (SLA) are marker autoantibodies for AIH. 52% of AIH patients are positive for ANA and/or SMA, 20% for SLA and 3% for LKM-1. These antibodies are of diagnostic value for AIH but the only autoantibodies highly specific for AIH are SLA. ANA/SMA also occur in 10-15% of patients with viral hepatitis and other immune-mediated diseases. LKM-1 are also associated with hepatitis C. Three types of LKM antibodies can be distinguished according to the target antigens. LKM-1 antibodies are directed against cytochrome p450 IID6, a 50 kDa cytoplasmic protein found in hepatocytes and renal proximal tubular cells. LKM-2 antibodies are associated with ticrynafen (tienilic acid) –induced hepatitis. The target antigen is cytochrome p450 IIC9, a cytochrome p450 isoenzyme that catalyzes the metabolic oxidation of the drug. LKM-3 antibodies are associated with chronic hepatitis D. The target antigen is UDP-1 glucoronosyl transferase. LKM-1 associated AIH predominantly occurs in girls between 2 and 14 years of age, thus determination of LKM-1 is very important in pediatrics.

Standard Curve

Do NOT use this example for interpreting sample's result.