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Description: EndoCab IgM, Human, ELISA

The EndoCab® IgM 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 compared to 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 endotoxin-core 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 IgM (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 endotoxin-core antibody 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 controled 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-IgM
Product type	Assays
Quantity	1 x 96 det.
Standard range	0.05 - 3.5 MMU/ml
Detection level	0.05 MMU/ml
Working volume	100 µl/well
Species	Human
Application	The EndoCab® IgA ELISA kit is to be used for the in vitro quantitative determination of endotoxin-core antibodies in serum, plasma and cell culture supernatant.
Disease	Gastroenterology
Principle	The EndoCab® IgA 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 a 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. Anti-endotoxin-core antibodies are captured by the solid phase antigen. Peroxidase conjugated anti-human IgA antibody will bind to captured endotoxin-core antibodies. The peroxidase conjugated antibody will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic 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 Endocab® IgA standards (log). The endotoxin-core concentration of samples, which are run concurrently with the standards, can be determined from the 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 6. Redd, A et al; Microbial translocation, the innate cytokine response and HIV-1 disease progression in Africa. PNAS 2009, 106: 6718 7. Boleij, A et al; Increased Exposure to Bacterial Antigen RpL7/L12 in Early Stage Colorectal Cancer Patients. Cancer 2010, 4014 8. Pasternak, B et al; Lipopolysaccharide Exposure Is Linked to Activation of the Acute Phase Response and Growth Failure in Pediatric Crohn's Disease and Murine Colitis. Inflamm Bowel Dis 2010, 16: 856 9. Bukh, A et al; Endotoxemia Is Associated with Altered Innate and Adaptive Immune Responses in Untreated HIV-1 Infected Individuals. PlosOne 2011, 6: e21275 10. Mondal, D et al; Contribution of Enteric Infection, Altered Intestinal Barrier Function, and Maternal Malnutrition to Infant Malnutrition in Bangladesh. CID 2011 11. Dello, S et al; Total Intermittent Pringle Maneuver during Liver Resection Can Induce Intestinal Epithelial Cell Damage and Endotoxemia. PlosOne 2012, 7: e30539