Product Name:	Cobalt Assay Kit (BA0213)
SKU:	BA0213
Detection Method:	Colorimetric (OD 485 nm)
Detection Range:	1.4 - 200 uM cobalt
Sample Type:	Aqueous samples
Species Reactivity:	All
Assay Time:	Within 10 minutes
Kit Size:	100 Assays
Equipment Required:	Microplate reader
Storage:	Shipped at room temperature. Store Detection Reagent at -20 degrees C.
Shelf Life:	6 months after receipt
Shipping:	Room Temperature

Cobalt is an essential micronutrient for all multicellular organisms as the active centre of cobalamins, such as vitamin B-12, which is essential for plants and animals. Cobalt is also a micronutrient for bacteria, algae and fungi. Despite being an important micronutrient, excess cobalt in the environment can be deleterious to life, leading to cardiomyopathy in humans and necrosis in plants. Simple, direct and automation-ready procedures for measuring cobalt concentrations find wide applications in research and environmental monitoring. This cobalt assay kit is designed to measure total cobalt directly in aqueous samples without any pretreatment. The intensity of the colour, measured at 485 nm, is directly proportional to the cobalt concentration in the sample.

Sensitive and accurate. Linear detection range 1.4 uM to 200 uM cobalt in 96-well plate assay.
Simple and high-throughput. The procedure involves addition of a single working reagent with immediate colour development. Can be readily automated as a high-throughput assay for thousands of samples per day.
Improved reagent stability and versatility. The optimised formulation has greatly enhanced reagent stability. Cuvette or 96-well plate assay.

Direct assays of cobalt in aqueous samples.

Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Step	Procedure
1	For the 96-well plate procedure, directly transfer the 200 uM Cobalt Standards into a clear flat-bottom 96-well plate to give 200, 120, 60 and 0 uM (90 uL total per well) as described in the standard table.
2	Transfer 90 uL of each sample into a clear flat-bottom 96-well plate.
3	Add 10 uL of Detection Reagent to each well and tap the plate to mix.
4	Read the optical density at 485 nm within 10 minutes at room temperature.
5	For the cuvette procedure, prepare standards with appropriate volumes for the cuvettes, set up labelled centrifuge tubes for standards and samples, add Detection Reagent to all tubes at a ratio of 1:10, mix by vortexing, transfer to cuvettes and read the OD at 485 nm within 10 minutes.

Subtract the OD of the 0 uM cobalt standard from all other standard OD values and plot the OD against standard cobalt concentrations. Determine the slope using linear regression fitting. The cobalt concentration of the sample is calculated as: [Co2+] = (OD_SAMPLE - OD_BLANK) / Slope (uM), where OD_BLANK is the OD value of the water blank, or of the sample blank if a sample blank is used.