Journal: Bio-protocol

Article Title: Quantification of 20-Hydroxyecdysone From Drosophila Larvae at the Onset of Metamorphosis

doi: 10.21769/BioProtoc.5831

Figure Lengend Snippet: Preparation of different standard solutions (Std1–7 and B0) of 20E for ELISA using 1× assay buffer as diluent.

Article Snippet: ELISA plate shaker (Neuation, model: iSHAK TS4 NXT)

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Bio-protocol

Article Title: Quantification of 20-Hydroxyecdysone From Drosophila Larvae at the Onset of Metamorphosis

doi: 10.21769/BioProtoc.5831

Figure Lengend Snippet: Reagent and buffer volumes for competitive ELISA setup

Article Snippet: ELISA plate shaker (Neuation, model: iSHAK TS4 NXT)

Techniques: Competitive ELISA, Enzyme Immunoassay

Journal: Bio-protocol

Article Title: Quantification of 20-Hydroxyecdysone From Drosophila Larvae at the Onset of Metamorphosis

doi: 10.21769/BioProtoc.5831

Figure Lengend Snippet: The principle of 20E estimation by this method relies on competitive ELISA. The presence of unlabeled 20E in samples will compete with 20E-HRP for binding to anti-20E antibodies and enzymatic reaction-dependent color generation. (A) Blank represents anti-rabbit antibody-coated wells without any buffers or reagents (no binding, no reaction, and no color). (B) NSB indicates minimal or nonspecific background binding (nonspecific adherence of 20E-HRP to the substratum and thus background reactivity and color generation). (C) B0 represents the maximum binding of 20E-HRP in the absence of free 20E (no competition, maximum binding, maximum enzymatic activity, highest color generation). (D) Standard demonstrates the inverse relationship between 20E-HRP binding and the concentration of free 20E (available 20E amount-dependent competition, curtailed enzyme activity, reduced color generation).

Article Snippet: ELISA plate shaker (Neuation, model: iSHAK TS4 NXT)

Techniques: Competitive ELISA, Binding Assay, Nonspecific Binding, Saturation Binding, Activity Assay, Concentration Assay