QuantiChrom™ Boron Assay Kit

Protocol SDS

Application

• For quantitative determination of boron in agricultural and environmental samples.

Key Features

• Fast and sensitive. Linear detection range: 0.05 to 10 µg/mL (0.05 – 10 ppm) boron with 100 µL sample (96-well).
• Convenient. The procedure involves adding a single working reagent.
• High-throughput. “Add-mix-read” type assay. Can be readily automated as a high-throughput 96-well or 384-well plate assay for thousands of samples per day.

Method

• OD420nm

Samples

• Water, plant tisse, soil samples, and antibody conjugation solutions

Species

• All

Procedure

• 40 min

Size

• 100 tests

Detection Limit

• 0.05 µg/mL or 0.05 ppm

Shelf Life

• 12 months

More Details

BORON is an essential micronutrient in plants and is involved in maintaining robust cell walls, cell membranes, and reproductive tissues. Although boron is common in the soil in its natural state as a borate mineral, the amount of boron available to plants is actually quite small. As a result, boron deficiency is the second most common micronutrient deficiency among crop plants. In order to keep plant boron levels in a healthy range, supplementation to the soil via fertilizers and additives is often required. If not regulated, a lack of or excess of boron may significantly lower crop yield. In the biotech industry, sodium borohydride is commonly used to conjugate antibodies and typically needs to be removed from the final product, especially for therapeutic antibodies.BioAssay Systems’ boron detection kit provides a convenient and reliable means to measure boron. In the assay borate complexes with azomethine-H to create a colored compound that can be measured at 420 nm. This assay can be used with a variety of samples and is simple, sensitive, and adaptable to high-throughput screening.

My sample has background absorbance at OD420nm; will this be an issue with the assay?

The calculation provided in the protocol accounts for sample background by subtracting the sample background blank.

After I make the working reagent, how long is it good for?

The working reagent should be made fresh and used within 15 minutes.

My samples have very high/low ODs, what is happening?

If your samples have very low ODs you may be below the kit detection limit. If your sample is from a soil or plant extraction, consider a more concentrated extraction to increase the amount of boron in the sample. If your samples have high ODs consider diluting them so they fall within the linear range of the standard curve.

Can I use this kit to determine the level of boron in my metal, glass, etc.?

This kit is optimized to detect boron in the form of borates and cannot be guaranteed to work with synthesized, and/or exotic boron compounds. However, samples may be treated to convert boron to borates.

How can I extract boron from a soil sample?

For dry weight analysis, first completely dry the soil to remove all water weight. Next, we recommend mixing 5 g of soil (sieved to remove rocks and debris) with 50 mL of 50 mM HCl for 30 min. Centrifuge the sample to pellet precipitates and use the supernatant for the assay. Note: if there are soil particles floating make sure to avoid them when pipetting; your sample should be completely free of particulate matter.

How can I extract boron from a plant sample?

For dry weight analysis, first completely dry the plant matter to remove all water weight. Next, we recommend burning 5 to 10 g of plant matter to ashes in a crucible at 500 degrees Fahrenheit for 3 hours in a drying oven. You may also use a blow torch to burn to ashes if a drying oven is not available. Mix all of the ashed plant material with 50 mL of 50 mM HCl for 30 min. Centrifuge the sample to pellet the ash and use the supernatant for the assay. Note: if there are ash particles floating make sure to avoid them when pipetting; your sample should be completely free of particulate matter.

My sample is in an acidic/basic solution, can I use it for the assay?

If samples are strongly acidic or basic they should be adjusted to a pH of 5-7. Weak or dilute acids and bases do not need to be pH adjusted as the Working Reagent has sufficient buffering capacity.

Why do you subtract the water OD from the background OD in the formula?

In the [Boron] equation the OD_BLANK is subtracted from the OD_SAMPLE to remove any contribution from the reagent, plate, and reaction volume from the OD of the sample. OD_H2O is subtracted from the OD_{SAMPLE BG} so that only the background absorbance of the sample is further subtracted from the OD_SAMPLE or, in other words, to make sure that the plate and reaction volume contributions to the OD_SAMPLE are not subtracted out twice.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.