Description: EndoCab, Human, ELISA kit

The EndoCab® ELISA has been developed for determination of endotoxin core antibodies in human plasma or serum in patients or healthy individuals. Several studies show a consistent drop in postoperative levels of circulating anti-endotoxin core antibodies from the preoperative value. This drop has been interpreted as consumption of antibodies to endotoxin by systemic release of endotoxin. A hypothesis is that if the patients pre-operative EndoCab® level is low, even moderately low, patients may not be able to cope with the efflux of endotoxin, which may have mild to severe clinical consequences. The assay is of interest for various experimental conditions ranging from in vitro LPS neutralization by plasma components to in vivo studies on kinetics of antibodies to endotoxin in health and diseases. The EndoCab® standard median-units IgG, IgM and IgA (MU) are arbitrary and are based on medians of ranges for 1000 healthy adults in a particular locality. It has not been established whether normal EndoCab® values vary by region, culture or race. Users should establish appropriate local statistical controls for their studies. It is advised that studies of any patient group should always be controlled by studies of appropriately selected contrasting clinical groups and/or healthy individuals recruited locally. EndoCab® is a registered trademark. Used under license from the Scottish National Blood Transfusion Service.

Specifications

Catalog number HK504
Product type Assays
Quantity 1 x 96 det.
Standard range Standard curve of IgG from 0.13-8 GMU/­ml. Standard curve of IgM from 0.05-3.5 MMU/ml. Standard curve of IgA from 0.16-10 AMU/ml.
Detection level Minimum detection level IgG EndoCab® antibodies is 0.13 GMU/ml. Minimum detection level IgM EndoCab® antibodies is 0.05 MMU/ml. Minimum detection level IgA EndoCab® antibodies is 0.16 AMU/ml.
Working volume 100 µl/well
Species Human
Application The EndoCab® ELISA is intended for the quantitative measurement of endotoxin-core antibodies (EndoCab®) in cell culture medium, plasma or serum.
Disease Gastroenterology
Principle The EndoCab® ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 2½ hours. The efficient format of 1 plate with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are incubated in microtiter wells coated with equimolar amounts of endotoxin rough-lipopolysaccharides from four Gram-negative bacterial species, each comprised of a complete inner core, but lacking complete outer core or O-specific polysaccharide chain. These endotoxins, complexed with polymyxin B, constitute the solid phase antigen. During this incubation anti-endotoxin-core antibodies are captured by the solid phase antigen. Unbound material present in the sample is removed by wash­ing. Peroxidase conjugated anti-human Ig(G or M or A) tracer antibody is added to the wells. If EndoCab antibodies were present in the sample, tracer antibodies will bind to the captured EndoCab antibodies. Excess tracer is removed by washing and substra­te, tetramethylben­zidine (TMB), is added to the wells. Color develops proportionally to the amount of anti endotoxin core antibodies present in the sample. The enzyme reaction is stopped by the addition of citric acid. The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the LBP standards (log). The human EndoCab® antibody concentration of samples, which are run concurrently with the human LBP standards, can be determined from the human LBP standard curve.
Storage and Stability Product should be stored at 4 °C. Under recommended storage conditions, product is stable for one year.
Precautions For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
References 1. Barclay, GR; Endogenous endotoxin-core antibody (EndoCab) as a marker of endotoxin exposure and a prognostic indicator: a review. Prog Clin Biol Res 1995, 392: 263
2. Allan, E et al; Anti-bacteroides lipopolysaccharide IgG levels in healthy adults and sepsis patients. Immunol Med Microbiol 1995, 11: 115
3. Barclay, GR, Antibodies to endotoxin in health and disease. Rev Med Midrobiol 1990, 1:133
4. Rahman, S et al; Intestinal hypoperfusion contributes to gut barrier failure in severe acute pancreatitis. J Gastrointest Surg 2003, 7: 26
5. Eckerwall, G et al; Early nasogastric feeding in predicted severe acute pancreatitis: A clinical, randomized study. Ann Surg 2006, 244: 959