Anti-Prothrombin IgM ELISA Kit

Sample

serum

Species Reactivity

Human

Detection Method

iELISA

Intended Use

Prothrombin-M is a solid phase enzyme immunoassay employing native human prothrombin (Factor II) for the quantitative detection of antibodies against prothrombin in human serum. The assay is a tool in the research of the antiphospholipid syndrome.

Contents of Kit

To be reconstituted:
1. 5× Sample Buffer 1 vial, 20 ml - 5× concentrated (capped white: yellow solution). Containing: Tris, NaCl, BSA, sodium azide (preservative)
2. 50× Wash Buffer 1 vial, 20 ml - 50× concentrated (capped white: green solution). Containing: Tris, NaCl, Tween, sodium azide (preservative)
Ready to use:
1. Negative Control 1 vial, 1.5 ml (capped green: yellow solution). Containing: Human serum (diluted), sodium azide (preservative)
2. Positive Control 1 vial, 1.5 ml (capped red: yellow solution). Containing: Human serum (diluted), sodium azide (preservative)
3. Calibrators 6 vials, 1.5 ml each 0, 3, 10, 30, 100, 300 U/ml. (color increasing with concentration : yellow solution). Containing: Human serum (diluted), sodium azide (preservative)
4. Conjugate 1 vial,12 ml IgM (capped green: green solution). Containing: Anti-human immunoglobulins conjugated to horseradish peroxidase
5. TMB Substrate 1 vial, 12 ml (capped black). Containing: Stabilized TMB/H₂O₂
6. Stop Solution 1 vial, 15 ml (capped white: colorless solution). Containing: 1M Hydrochloric Acid
7. Microtiterplate 12×8 well strips with breakaway microwells. Coating see paragraph 1

Storage

At 2-8°C use original vials, only.
Store all reagents and the microplate at 2-8°C, in their original containers. Once prepared, reconstituted solutions are stable for 1 month at 4°C, 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.

Detection Range

0-300 U/ml

Sensitivity

1.0 U/ml

General Description

Human Factor II (prothrombin) is a plasme zymogen with a molecular weight of 72 kDa. It assembles with the activated forms of Factor V, Factor X and phospholipid to form a catalytic unit known as the prothrombinase complex. In the presence of calcium ions, the complex cleaves the membrane-associated prothrombin into thrombin, which is then released into the soluble phase.
Antibodies against prothrombin belong to the group of antiphospholipid antibodies speciffc for negatively charged phospholipids, components of biological membranes, and prothrombin. The presence of anti-phospholipid antibodies has been associated with the clinical features of the so-called antiphospholipid syndrome (APS). Many studies have shown a correlation between these autoantibodies and an enhanced incidence of thrombosis, thrombocytopenia and habitual abortions (as a consequence of placental infarct). The exact mechanism by which pathogenic anti-phospholipid antibodies induce thrombosis is not yet revealed fully.
The occurrence of anti-phospholipid antibodies in patients with SLE and related diseases is typical for a secondary anti-phospholipid syndrome (APS). In contrast, anti-phospholipid antibodies in patients with no other autoimmune diseases characterize the primary anti-phospholipid syndrome (APS).

Standard Curve

Example of a standard curve
We recommend pipetting calibrators in parallel for each run.

Medical laboratories might perform an in-house Quality Control by using own controls and/or internal pooled sera, as foreseen by EU regulations.