Indirect ELISA assays were used to characterize the degree of biotinylation of BDNF. All ELISA reactions were performed at room temperature unless otherwise noted. Briefly, 96-well plates (NUNC, Thermo Scientific, Rochester, NY) were coated with 75 ng/ml anti-human BDNF antibodies (R&D systems, Minneapolis, MN; Cat# MAB848) in 25 mM carbonate buffer pH 9.7 overnight at 4°C. Next, plates were blocked with ELISA blocking solution (Pierce; Cat# N502) for 1 h, incubated with either biotinylated or unmodified BDNF at different concentrations (0.625–40 ng/ml) for 2 h, followed by avidin-HRP (Pierce; Cat# N100) at 1∶10000 dilution for 1 h, and developed with K-blue MAX TMB substrate (Tetramethylbenzidine; Neogen, Lansing, MI). The reaction was stopped with 1 N HCl, and absorbance was measured at 450 nm using a microplate reader (Infinite 200 PRO, Tecan, Männerdorf, Switzerland).
Anti human bdnf antibodies
Manufactured by R&D Systems
Anti-human BDNF antibodies are laboratory reagents used for the detection and quantification of brain-derived neurotrophic factor (BDNF) in human samples. These antibodies specifically bind to BDNF, allowing researchers to measure its levels in various biological matrices, such as cell lysates, tissue homogenates, and biological fluids.
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Lab products found in correlation
Infinite 200 pro, by Tecan (1 mentions)
Absorbent pad, by Cytiva (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Sample pad, by Cytiva (1 mentions)
Tris base, by Merck Group (1 mentions)
Tris hydrochloride, by Merck Group (1 mentions)
Varioskan lux multimode, by Thermo Fisher Scientific (1 mentions)
Whatman, by Cytiva (1 mentions)
2 protocols using anti human bdnf antibodies
1
Quantification of BDNF Biotinylation by ELISA
2
Lateral Flow BDNF Assay Development
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A human BDNF ELISA kit, recombinant anti-mouse IgG2a antibodies, and recombinant human BDNF protein were obtained from Abcam. Albumin, bovine serum albumin, calcium chloride, glucose, gold nanoparticles (40 nm), magnesium chloride, potassium chloride, sucrose, Tween 20, sodium chloride, Tris-hydrochloride and Tris base were obtained from Sigma-Aldrich. Two different anti-Human BDNF antibodies were purchased from R&D Systems. All components of the lateral flow test strip, including the backing, sample pad, conjugate pad, reacting membrane and absorbent pad were obtained from Whatman. The absorbance spectrum was measured through a microplate reader (Varioskan LUX Multimode, ThermoFisher). The test line and control line were dispensed using a syringe pump (KDS-200-CE, KD Scientific Inc.). The readout box was produced using a 3D printer in Chemical Engineering Workshop. The images were captured using an iPhone X.
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