Name
EndoCab, Human, ELISA kit
Catalog nr
HK504
(lot number and expiry date are indicated on the label)
Short description
The EndoCab ® ELISA has been developed for determination of endotoxin core antibodies (EndoCab ® ) 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...
Size
1 x 96 det.
Application
Manual
Description
The EndoCab® ELISA has been developed for determination of endotoxin core antibodies (EndoCab®) 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.
Application
The EndoCab® ELISA is intended for the quantitative measurement of endotoxin-core antibodies (EndoCab®) in cell culture medium, plasma or serum. In serum or plasma samples IgG can be measured accurately if serum or plasma samples are diluted at least 200 times, IgM if samples are diluted at least 100 times and IgA if samples are diluted at least 50 times. Most reliable results are obtained if EDTA plasma is used. The EndoCab® ELISA kit can be used for different species, like rhesus monkey, rat and mouse. However, when using other samples then human, the standard and tracer have to be replaced by a pool plasma of the used species and the right corresponding HRPO labeled antibody (not included).
Features
- Minimum detection level IgG EndoCab® antibodies is 0.125 GMU/ml.
- Standard curve of IgG from 0.125‑8 GMU/ml.
- Minimum detection level IgM EndoCab® antibodies is 0.055 MMU/ml.
- Standard curve of IgM from 0.055-3.5 MMU/ml.
- Minimum detection level IgA EndoCab® antibodies is 0.156 AMU/ml.
- Standard curve of IgA from 0.156-10 AMU/ml.
- Working volume of 100 µl/well.
Typical standard curve
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 at least six months. After reconstitution the reagents are stable for 1 month if stored at 2-8°C.
References
- 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
- Allan E et al., Anti-bacteroides lipopolysaccharide IgG levels in healthy adults and sepsis patients. Immunol Med Microbiol, 1995, 11: 5
- Barclay GR, Antibodies to endotoxin in health and disease. Rev Med Midrobiol, 1990, 1: 133
- Reyes, H et al; Is a leaky gut involved in the pathogenesis of intrahepatic cholestatis of pregnancy? Hepatology 2006, 43:715
- Braun, J et al; Barrier function of the gut and multiple organ dysfunction after cardiac surgery. J of Int Med Res 2007, 35:72
- Boleij A et. al, Increased exposure to bacterial antigen RpL7/L12 in early stage colorectal cancer patients. Cancer 2010, 116:4014
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.
Also available
References
- 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
- Allan E et al., Anti-bacteroides lipopolysaccharide IgG levels in healthy adults and sepsis patients. Immunol Med Microbiol, 1995, 11: 5
- Barclay GR, Antibodies to endotoxin in health and disease. Rev Med Midrobiol, 1990, 1: 133
- Reyes, H et al; Is a leaky gut involved in the pathogenesis of intrahepatic cholestatis of pregnancy? Hepatology 2006, 43:715
- Braun, J et al; Barrier function of the gut and multiple organ dysfunction after cardiac surgery. J of Int Med Res 2007, 35:72
- Boleij A, Increased exposure to bacterial antigen RpL7/L12 in early stage colorectal cancer. Cancer 2010, 116:4014
MSDS
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