China
Africa
Explore chapters and articles related to this topic
Bioanalytical Methods
Published in Jerome Greyson, Carbon, Nitrogen, and Sulfur Pollutants and Their Determination in Air and Water, 2020
Antibodies are produced in the blood stream of an animal by repeatedly injecting it with antigen over a period of time. The repeated injections stimulate the animal to produce a high concentration (or titre) of antibody, which is then harvested by drawing aliquots of the animal’s blood. Because of the high specifity antibodies exhibit, using them generally requires little more purification than centrifuging the harvested blood to remove its red cells. The antibody-containing serum, i.e., the supernatant fluid remaining after the red blood cells are removed, is generally referred to as antiserum.
Ameliorative effect of vitamin E and selenium against bisphenol A-induced toxicity in spinal cord and submandibular salivary glands of adult male albino rats
Published in International Journal of Environmental Health Research, 2023
Dina W. Bashir, Yasmine H. Ahmed, Mohamed A. El-Sakhawy
For the detection of astrocyte protein, the SC was dissected, removed, fixed, and embedded in paraffin (Merck 1109). Paraffin sections (5 μm) were mounted onto glass slides coated with poly-L-lysine. To remove the paraffin wax, sections were treated in xylene and then acetone; 10 min in each. After a short rinse (1–5 min) in two changes of phosphate-buffered saline (PBS, pH 7.4), sections were treated for 30 min at room temperature with hydrogen peroxide and in methanol to exhaust endogenous peroxidase activity. The following steps all included two PBS rinses interposed between the change of immune reagents under continuous shaking for 5 min. To suppress background staining, sections were incubated in a wet chamber for 20 min at room temperature in normal goat serum. Incubation with a 1:500 dilution of polyclonal rabbit anti-GFAP antiserum (Dakopatts, Glostrup, Denmark) in PBS was carried out at 4°C for 24 h and eventually a second antibody (biotinylated mouse anti-rabbit immunoglobulin) for 30 min (the avidin-biotin complex). The method used was outlined by Stoltenburg et al. (1996).
Production of polyclonal antibody against human Neuritin and its application of immunodetection
Published in Preparative Biochemistry and Biotechnology, 2019
Na Wang, Yu Wei, Wen Zhang, Xingyi Li, Jingling Zhu, Liya Shan, Chunyan Liu, Wumei Yuan, Jin Huang
Antiserum was obtained by direct injection of protein[13,14] and the rhNeuritin was used to immunize Sprague-Dawley rats to prepare the polyclonal antibody. A total of three immunizations were administered within 35 days. The antiserum was collected and purified by immunoaffinity chromatography and showed high sensitivity, with titers as high as 1:16,000 as measured by indirect ELISA. The antibody recognized both exogenous and endogenous Neuritin as assessed by immunoblotting. However, affinity of the antibody to endogenous Neuritin was higher than exogenous Neuritin. Therefore, we investigated whether the antibody could also be used in other immunoassays. The antibody was used in immunofluorescence and successfully bound to Neuritin localized within lipid rafts in the cell membrane.[10]
Cloning, large-scale production and characterization of fusion protein (P-TUFT-ALT-2) of Brugian abundant larval transcript-2 with tuftsin in Pichia pastoris
Published in Preparative Biochemistry and Biotechnology, 2018
Rajkumar Paul, Selvarajan Karthik, Ponnusamy Vimalraj, Sankaranarayanan Meenakshisundaram, Perumal Kaliraj
Antibody level was tested against E-ALT-2 and P-TUFT-ALT-2 by indirect ELISA method. The 96-well microtitre plates (Nunc, Denmark) were coated with 100 µL of P-TUFT-ALT-2 (100 ng/well) prepared in 0.1 M bicarbonate buffer pH 9.6 and incubated overnight at 4 °C followed by blocking with 5% skimmed milk. The plates were washed and incubated with 100 µL of sera in a serial two-fold dilution (1:500–1,28,000) for 1 h 30 minutes at 37 °C. The same dilution was prepared for pre-immuned and alum immunized sera. After washes, the plates were incubated again with goat anti-mouse IgG-ALP conjugate (1:1000) for 45 min at 37 °C. The plates were washed and the color was developed using pNPP. Absorbance was read at 405 nm. The mean OD ±3SD of the pre-immune serum was taken as the cut-off value for determining antibody titer. The highest dilution of the antiserum that showed an OD value above the cut-off value was taken as the antibody titer.