Can the plant hormone ELISA kit simultaneously measure multiple indicators?

It is not recommended to pursue "measuring three indicators on one plate simultaneously"—different hormones may require different extraction solvents, dilution ratios, and competitive reaction conditions. You can purchase single-indicator strip-well kits or kits with three separate indicators, aliquot the same batch of extract, and follow each kit's respective instructions. If the sample volume is limited, prioritize single-well screening first, and use duplicate wells when necessary; three indicators at 50 μL each in single wells require at least 150 μL of reconstitution volume, so the post-extraction reconstitution volume must be calculated carefully. Some manufacturers offer multi-indicator plant hormone combinations, but you need to check whether they are suitable for Arabidopsis leaf tissue.

A few tens of milligrams of fresh weight is only enough for conducting very compact experiments. It is recommended to grind with liquid nitrogen and use 80% methanol or the manufacturer's recommended extraction solution for overnight extraction at 4 ℃. Centrifuge to obtain the supernatant, and then concentrate to near dryness with nitrogen blowing or vacuum. Finally, dissolve in a small volume, such as 80-120 μ L. After reconstitution, first perform one or two indicators and do not distribute them evenly; If retesting is not possible, sieve the single well first and retest for abnormal samples. The extraction and reconstitution volumes should be calculated in advance according to the minimum sample amount specified in the instructions.

Look for strip-well 48T or 96T formats, and clarify the minimum sample volume per well; 50 μL is ideal. Prioritize asking manufacturers whether they have validated data and extraction protocols for Arabidopsis/plant leaf samples, rather than general-purpose plant hormone kits. Regarding cost-effectiveness, domestic plant hormone-specific kits are usually within budget, but for targets such as GA3, cross-reactivity rates must be specifically confirmed. You can request the instruction manual and cross-reactivity table from the sales representative; do not make do with animal hormone ELISA kits.

Plant hormone small molecule ELISA often uses competitive methods to detect ABA, IAA, and GA3, which is theoretically feasible. However, GA3, a type of gibberellin antibody, is more commonly cross reactive and has lower specificity and sensitivity than LC-MS/MS. If it is only a relative comparison and trend screening between Arabidopsis mutants/treatments, ELISA can be used; If precise quantification is to be published or if there are doubts from reviewers, it is recommended to supplement LC-MS/MS later. The first time, it is necessary to perform spiked recovery and parallelism verification, especially for leaf extract matrices that are complex and may interfere with competitive reactions.