Product Name:	Detergent Assay Kit (BA0214)
SKU:	BA0214
Detection Method:	Colorimetric (OD 560 nm for Reagent A / OD 650 nm for Reagent B)
Detection Range:	Tween 80: 0.012-4 mM; Tween 20: 0.06-5 mM; Triton X-100: 0.23-12 mM; Brij L23/35: 0.09-5 mM; DTAC: 0.08-2 mM; SDS: 10-25 mM
Sample Type:	Purified protein samples, water and soil
Species Reactivity:	All
Assay Time:	Immediate (add-mix-read)
Kit Size:	100 Assays
Equipment Required:	Microplate reader
Storage:	Shipped at room temperature. Store contents at 4 degrees C.
Shelf Life:	12 months after receipt
Shipping:	Room Temperature

Detergents are surfactants that are amphiphilic, meaning that they are partly hydrophobic and partly hydrophilic. Detergents fall into three categories: anionic, cationic and non-ionic/zwitterionic. They are commonly used in household cleaning products, in gasoline as additives, and in biological reagents. Simple, direct and automation-ready procedures for measuring detergent concentration in biological samples are very desirable. This detergent assay kit is designed to measure detergent directly without any pretreatment. Above the critical micellar concentration, detergent forms micelles in solution that trap the colorimetric dye. The intensity of the colour is directly proportional to the detergent concentration in the sample.

Sensitive and accurate. Uses 20 uL sample. Detection range: Tween 80: 0.012 to 4 mM; Tween 20: 0.06 to 5 mM; Triton X-100: 0.23 to 12 mM; Brij L23/35: 0.09 to 5 mM; DTAC: 0.08 to 2 mM; SDS: 10 to 25 mM.
Simple and convenient. The procedure involves addition of a single reagent and measuring OD560nm or OD650nm.
Improved reagent stability and versatility. The optimised formulation has greatly enhanced reagent and signal stability. Cuvette or 96-well plate assay.

Protein purification: traces of detergent in purified protein samples.
Environment: detergent determination in water and soil.

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 Triton X-100 standard curve, dilute to 12 mM by mixing 45 uL of 100 mM Triton X-100 Standard with 330 uL of water, then dilute in water as described in the standard table; for the SDS standard curve, dilute to 25 mM by mixing 50 uL of 250 mM Standard with 450 uL of water and dilute as in the standard table.
2	Transfer 20 uL of diluted standards and samples into the wells of a clear flat-bottom plate; if the sample is coloured, transfer 20 uL of sample into two separate wells (one Sample and one Sample Blank).
3	Add 180 uL of Reagent A for cationic/nonionic detergents (e.g. Triton X-100), or 180 uL of Reagent B for anionic detergents (e.g. SDS), to all samples and standards; add 180 uL of deionised water to Sample Blank wells. Tap the plate to mix.
4	Read the optical density at 560 nm for Reagent A or 650 nm for Reagent B.
5	To construct a standard curve for other detergents, prepare at least eight dilutions above the critical micellar concentration plus a no-detergent water blank, select Reagent A for cationic/nonionic or Reagent B for anionic detergents, transfer 20 uL of standards into wells, add 180 uL of the appropriate reagent, read the OD, subtract the water blank OD and plot delta-OD against concentration; detergent standard curves are often nonlinear.

Subtract the blank OD (water) from the standard OD values and plot the OD against standard concentrations. Use the standard curve to determine the sample detergent concentration. If the calculated detergent is above the standard curve, dilute the sample in water and rerun the assay. Conversions: 1 mM Triton X-100 equals 0.0625% or 625 ppm; 1 mM SDS equals 0.0288% or 288 ppm.

Component	Quantity	Storage
Reagent A (for Cationic/Nonionic Detergents, e.g. Tween, Brij, Triton)	20 mL	4 degrees C
100 mM Triton X-100 Standard	100 uL	4 degrees C
Reagent B (for Anionic Detergents, e.g. SDS)	20 mL	4 degrees C
250 mM SDS Standard	100 uL	4 degrees C