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|Cell culture supernatant||120||106% - 133%|
|Milk||113||100% - 126%|
|Urine||87||80% - 93%|
|Serum||91||79% - 106%|
|Cell culture media||110||97% - 122%|
|Heparin Plasma||106||97% - 121%|
|EDTA Plasma||100||102% - 110%|
|Citrate Plasma||107||79% - 106%|
Antithrombin III in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of human Antithrombin III protein in serum, plasma, urine, milk, and cell culture supernatant samples.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Antithrombin-III (ATIII), also known as Serpin C1, is a 58 kDa extracellular glycoprotein belonging to the Serpin serine protease inhibitor superfamily. Antithrombin III functions primarily as a blood anticoagulant but also modulates inflammatory cytokine expression. Human Antithrombin-III is synthesized in the liver and is secreted as a 432 amino acid (aa) mature protein. Most circulating Antithrombin-III (90-95%) is the fully glycosylated α-isoform while the partially glycosylated β-isoform constitutes 5-10%. The biological activity of Antithrombin-III is inherently low and is enhanced up to 1000-fold in the presence of heparin or heparin-like glycosaminoglycans. The β-isoform of Antithrombin-III has a higher affinity for heparin. Mature human Antithrombin-III shares 90% sequence homology with mature mouse Antithrombin-III. Human Antithrombin-III inactivates thrombin and other blood coagulation factors. Human adult Antithrombin-III levels in plasma are relatively constant. However, reduction of Antithrombin-III is observed in individuals with diabetes or sepsis. Neonates also exhibit low levels of Serpin C1 in plasma.
|内容||1 x 96 tests|
|10X Wash Buffer PT (ab206977)||1 x 20ml|
|50X Cell Extraction Enhancer Solution (ab193971)||1 x 1ml|
|5X Cell Extraction Buffer PTR (ab193970)||1 x 10ml|
|Antibody Diluent 5BC||1 x 6ml|
|Denaturant||1 x 500µl|
|Human Antithrombin III Capture Antibody 10X||1 x 600µl|
|Human Antithrombin III Detector Antibody 10X||1 x 600µl|
|Human Antithrombin III Lyophilized Recombinant Protein||1 x 2 vials|
|Plate Seals||1 unit|
|Sample Diluent NS||1 x 50ml|
|SimpleStep Pre-Coated 96-Well Microplate (ab206978)||1 unit|
|Stop Solution||1 x 12ml|
|TMB Substrate||1 x 12ml|
Our Abpromise guarantee covers the use of ab222507 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent concentration.|
Background-subtracted data values (mean +/- SD) are graphed.
The concentrations of Antithrombin III were measured in duplicates, interpolated from the Antithrombin III standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:4,000 plasma (citrate) 1:8,000, plasma (EDTA) 1:4,000, and plasma (heparin) 1:4,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Antithrombin III concentration was determined to be 157.69 µg/mL in neat serum, 279.95 µg/mL in neat plasma (citrate), 160.27 µg/mL in neat plasma (EDTA), and 224.88 µg/mL in neat plasma (heparin).
The concentrations of Antithrombin III were measured in duplicates, interpolated from the Antithrombin III standard curves and corrected for sample dilution. Undiluted samples are as follows: milk 3.13%, Hep G2 supernatant 6.25%, and urine 6.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Antithrombin III concentration was determined to be 810.16 ng/mL in neat milk, 218.86 ng/mL in neat Hep G2 supernatant, and 618.60 ng/mL in neat spiked urine.
Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Antithrombin III concentration was determined to be 77.20 µg/mL with a range of 34.09 – 121.12 µg/mL.
ab222507 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"