Human alpha1-Antitrypsin ELISA Kit
alpha1-Antitrypsin ELISA
Reactivity
Human
Synonyms
alpha1-Antitrypsin; alpha1-Antitrypsin ELISA; alpha1-Antitrypsin elisa kit
Reactivity
Human
Specificity
No cross reactivity with other plasma proteins in stool was observed. No cross reactivity with alpha-1-antitrypsin in mouse serum was observed.
Sequence Length
409
Samples
Stool
Assay Type
Sandwich
Sensitivity
1.8 mg/dl.
Intra-assay Precision
The precision (intra-assay variation) of the alpha1-antitrypsin ELISA test was calculated from 20 replicate determinations on each one of two samples
Inter-assay Precision
The total precision (inter-assay variation) of the alpha1-antitrypsin ELI-SA test was calculated from data on 2 samples obtained in 20 different assays by three technicians on two different lots of reagents over a period of three months.
Preparation and Storage
2 - 8 degree C.
Related Product Information for alpha1-Antitrypsin elisa kit
Intended Uses: The described Enzyme-Linked-Immuno-Sorbent-Assay (ELISA) is intended for the determination of alpha1-antitrypsin in stool. For research use only.
Principle of the Assay: The assay utilizes the sandwich technique with two selected polyclonal antibodies that bind to human alpha1-antitrypsin. Standards, controls and prediluted samples which are assayed for human alpha1-antitrypsin are added into the wells of a micro plate coated with a high affine polyclonal anti-human alpha1-antitrypsin antibody. During the first incubation step, alpha1-antitrypsin is bound by the immobilized antibody. Then a peroxidase-conjugated polyclonal anti-human alpha1-antitrypsin antibody is added into each microtiter well and a sandwich of capture antibody-human alpha1-antitrypsin-peroxidase-conjugate is formed. Tetramethylbenzidine is used as peroxidase substrate. Finally, an acidic stop solution is added to terminate the reaction. The colour changes from blue to yellow. The intensity of the yellow colour is directly proportional to the concentration of alpha1-antitrypsin. A dose response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated, using the values obtained from the standard. alpha1-antitrypsin present in the samples is determined directly from this curve!!Background/Introduction: Intestinal protein loss is a serious consequence of various systemic or local gastrointestinal diseases (e. g. allergies, chronic inflammation, malignancies). These pathologies damage the mucosal integrity and/or cause lymphostasis, thereby leading to an increased transfer of plasma proteins into the bowel lumen. Subsequently, hypoproteinemia accompanied with edema may develop. This condition is identified by exclusion of other sources of protein loss and by proof of an elevated alpha1-antitrypsin concentration in stool. In serum, alpha1-antitrypsin represents the majority of serine protease inhibitors and protects tissues from protease damages during inflammation. The protein is synthesized primarily in the liver but also to a small extent in intestinal macrophages, monocytes, and intestinal epithelial cells. Since alpha1-antitrypsin is relatively resistant against enzymatic digestion, the secreted amount in stool reflects the internal concentration of the protein. An elevated alpha1-antitrypsin stool concentration is therefore a widely recognized marker for intestinal protein loss and for an increased mucosal permeability. In clinical routine, the alpha1-antitrypsin clearance (ratio of the alpha1-antitrypsin ELISA values of stool and serum samples) has been established along with the sole determination of the 24h alpha1-antitrypsin secretion in stool. Thus the group of J. S. Fordtran reports that the sole determination of the alpha1-antitrypsin concentration in stool yielded false positive or false negative results in 21 % of the subjects compared to the alpha1-antitrypsin clearance measurement (Strygler et al. 1990). The analytical quality of alpha1-antitrypsin ELISA surpasses by far the conventional radial immunodiffusion (RID) technique in the determination of serum, stool and tissue culture supernatants. In direct comparison, the concentrations measured with the ELISA were approximately 30 % above the corresponding RID levels. Cell culture supernatants of an intestinal cell line as well as fecal samples of lymphostasis subjects yielded negative results with RID. Our ELISA could detect alpha1-antitrypsin in all of these samples, in some of them even in very high concentrations. These results clearly prove that the alpha1-antitrypsin ELISA is far more sensitive than the conventional method and that it recognizes not only hepatic but also enteral alpha1-antitrypsin. The discrepancy of both methods and hence the superiority of the ELISA to RID is especially striking in the analysis of extremely high enteral protein losses. The combination of two specific antibodies in our alpha1-antitrypsin ELISA widely excludes the possibility of false negative results thereby enabling a reliable enteral protein loss.
Principle of the Assay: The assay utilizes the sandwich technique with two selected polyclonal antibodies that bind to human alpha1-antitrypsin. Standards, controls and prediluted samples which are assayed for human alpha1-antitrypsin are added into the wells of a micro plate coated with a high affine polyclonal anti-human alpha1-antitrypsin antibody. During the first incubation step, alpha1-antitrypsin is bound by the immobilized antibody. Then a peroxidase-conjugated polyclonal anti-human alpha1-antitrypsin antibody is added into each microtiter well and a sandwich of capture antibody-human alpha1-antitrypsin-peroxidase-conjugate is formed. Tetramethylbenzidine is used as peroxidase substrate. Finally, an acidic stop solution is added to terminate the reaction. The colour changes from blue to yellow. The intensity of the yellow colour is directly proportional to the concentration of alpha1-antitrypsin. A dose response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated, using the values obtained from the standard. alpha1-antitrypsin present in the samples is determined directly from this curve!!Background/Introduction: Intestinal protein loss is a serious consequence of various systemic or local gastrointestinal diseases (e. g. allergies, chronic inflammation, malignancies). These pathologies damage the mucosal integrity and/or cause lymphostasis, thereby leading to an increased transfer of plasma proteins into the bowel lumen. Subsequently, hypoproteinemia accompanied with edema may develop. This condition is identified by exclusion of other sources of protein loss and by proof of an elevated alpha1-antitrypsin concentration in stool. In serum, alpha1-antitrypsin represents the majority of serine protease inhibitors and protects tissues from protease damages during inflammation. The protein is synthesized primarily in the liver but also to a small extent in intestinal macrophages, monocytes, and intestinal epithelial cells. Since alpha1-antitrypsin is relatively resistant against enzymatic digestion, the secreted amount in stool reflects the internal concentration of the protein. An elevated alpha1-antitrypsin stool concentration is therefore a widely recognized marker for intestinal protein loss and for an increased mucosal permeability. In clinical routine, the alpha1-antitrypsin clearance (ratio of the alpha1-antitrypsin ELISA values of stool and serum samples) has been established along with the sole determination of the 24h alpha1-antitrypsin secretion in stool. Thus the group of J. S. Fordtran reports that the sole determination of the alpha1-antitrypsin concentration in stool yielded false positive or false negative results in 21 % of the subjects compared to the alpha1-antitrypsin clearance measurement (Strygler et al. 1990). The analytical quality of alpha1-antitrypsin ELISA surpasses by far the conventional radial immunodiffusion (RID) technique in the determination of serum, stool and tissue culture supernatants. In direct comparison, the concentrations measured with the ELISA were approximately 30 % above the corresponding RID levels. Cell culture supernatants of an intestinal cell line as well as fecal samples of lymphostasis subjects yielded negative results with RID. Our ELISA could detect alpha1-antitrypsin in all of these samples, in some of them even in very high concentrations. These results clearly prove that the alpha1-antitrypsin ELISA is far more sensitive than the conventional method and that it recognizes not only hepatic but also enteral alpha1-antitrypsin. The discrepancy of both methods and hence the superiority of the ELISA to RID is especially striking in the analysis of extremely high enteral protein losses. The combination of two specific antibodies in our alpha1-antitrypsin ELISA widely excludes the possibility of false negative results thereby enabling a reliable enteral protein loss.
Product Categories/Family for alpha1-Antitrypsin elisa kit
NCBI and Uniprot Product Information
NCBI GI #
NCBI Official Full Name
alpha1 antitrypsin
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Product Notes
The Human alpha1-Antitrypsin (Catalog #AAA495094) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA495094 ELISA Kit recognizes Human alpha1-Antitrypsin. It is sometimes possible for the material contained within the vial of "alpha1-Antitrypsin, ELISA Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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