ELISA kits with colorimetric output were used to determine the plasma concentrations of PAI-1 (ab184863, Abcam), fibrinogen (ab241383, Abcam), plasminogen (ab108893, Abcam), tissue plasminogen activator (t-PA) (ab190812, Abcam), platelet factor 4 (PF4) (ab189573, Abcam), interleukin (IL) 1 receptor-like 1 (ST2) (ab254505, Abcam), von Willebrand factor (vWF) (ab223864, Abcam), and tryptase (EKU10581, Biomatik) according to the manufacturer’s instructions. The plates were read at 450 nm using an automated microplate reader (Biotek Epoch). For each subject, we report the mean value from at least two replicate determinations with a coefficient of variation (CV) lower than 20%. The mean intra-assay CV was lower than 10% for all biomarkers and the inter-assay CVs were 11% for PAI-1, 10% for fibrinogen, 7% for plasminogen, 8% for t-PA, 9% for PF4, 14% for ST2, 10% for vWF, and 12% for tryptase. The plasma concentrations of each biomarker were determined using a standard curve, constructed and fit using a 4P logistic regression equation, and the assay values were corrected by the appropriate dilution factor in each case.
Ab184863
Manufactured by Abcam
Sourced in United Kingdom
Ab184863 is a lab equipment product offered by Abcam. It is a core functional device designed for laboratory applications. No further details are available.
Automatically generated - may contain errors
Lab products found in correlation
Ab241383, by Abcam (1 mentions)
Ab189573, by Abcam (1 mentions)
Ab108893, by Abcam (1 mentions)
Ab223864, by Abcam (1 mentions)
Sample buffer, by Thermo Fisher Scientific (1 mentions)
Sample reducing agent, by Thermo Fisher Scientific (1 mentions)
P0448, by Agilent Technologies (1 mentions)
P0447, by Agilent Technologies (1 mentions)
Coomassie brilliant blue, by Fujifilm (1 mentions)
2 protocols using ab184863
1
Multiplex Biomarker Quantification in Plasma
2
Isolation and Characterization of Salivary SIgA
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We purified SIgA using an IgA purification kit (#20395; Thermo Fisher Scientific, Waltham, MA, USA) as described by the manufacturer. Purified saliva samples were added to the mixture of sample buffer (#NP0008; Thermo Fisher Scientific) and sample reducing agent (#NP0009; Thermo Fisher Scientific). The samples were heated for 5 min at 96°C and run on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel using a standard protocol. The gel was stained with Coomassie brilliant blue (#178–00551; FUJIFILM Wako Chemicals, Osaka, Japan). Antibodies specific for the heavy chain of IgA were used to determine whether the purified substance was IgA. We further confirmed that the detected IgA was the secretory type (SIgA) using an antibody specific to the secretory component. Western blotting was employed using the following primary antibodies: anti-human IgA rabbit monoclonal antibody (ab184863; Abcam Plc, Cambridge, UK; 1:500 dilution) and anti-human IgA SC mouse monoclonal antibody (ab3924; Abcam, 1:500 dilution). Horseradish peroxidase-conjugated anti-rabbit polyclonal antibody (#P0448; Dako, Glostrup, Denmark; 1:1000 dilution) or anti-mouse monoclonal antibody (#P0447; Dako; 1:1000 dilution) was used as the secondary antibody.
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