ELISA plates (Corning, Pittsburg, PA) were coated overnight with 2μg/mL of recombinant human HER2 Fc chimera (R&D Systems, Minneapolis, MN). The next day, different amounts of trastuzumab (1ng-5μg/mL) and culture supernatant were applied and incubated for 2hr. Non-specific binding was eliminated by vigorous washes with TBS-Tween (Boston Bioproducts, MA). The plates were then incubated with secondary antibody, anti-human IgG (Fab specific)-alkaline phosphatase conjugate (Sigma, St Louis, MO) for 2hr. After washing with TBS-Tween, the plates were incubated with pNPP substrate (Sigma, St. Louis, MO) for 5-15 minutes and the optical density was measured at 405nM.
Anti human igg fab specific alkaline phosphatase conjugate
Manufactured by Merck Group
The Anti-human IgG (Fab specific)-alkaline phosphatase conjugate is a laboratory reagent used for the detection and quantification of human immunoglobulin G (IgG) in various immunoassay applications. It consists of an anti-human IgG antibody fragment (Fab) that is conjugated to the enzyme alkaline phosphatase. This conjugate is designed to specifically bind to the Fab region of human IgG, enabling the sensitive and specific detection of IgG in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Recombinant human her2 fc chimera, by R&D Systems (2 mentions)
Elisa plate, by Corning (2 mentions)
Pnpp substrate, by Merck Group (1 mentions)
Tbs tween, by Boston BioProducts (1 mentions)
Tbs tween, by Merck Group (1 mentions)
Para nitro phenyl phosphate pnpp substrate, by Merck Group (1 mentions)
2 protocols using anti human igg fab specific alkaline phosphatase conjugate
1
Quantifying HER2 Binding Kinetics
2
ELISA-based HER2 Antibody Quantification
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Enzyme-linked immunosorbent assay (ELISA) plates (Corning) were coated overnight with 2 μg/mL recombinant human HER2 Fc chimera (R&D Systems). The following day, 100 µl culture supernatant of either vector control or LM008–HER2Ab cells were applied and incubated for 2 h at RT. Increasing concentrations of trastuzumab (0 to 200 ng/mL) were also incubated and used as a positive control. Nonspecific binding was eliminated by vigorous washes with Tris-buffered saline (TBS)-Tween (Boston Bioproducts). The plates were then incubated with secondary antibody, anti-human IgG (Fab specific)-alkaline phosphatase conjugate (Sigma) for 2 h. After washing with TBS-Tween, plates were then incubated with para-Nitrophenyl Phosphate (pNPP) substrate (Sigma) for 5 to 15 min, and the optical density was measured at 405 nM.
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