Formaldehyde Detection Kit

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Antigen
Minimum Detection Limit
0.848 μM
Application
Biochemical Assay (BCA)
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Purpose The DetectX® Formaldehyde kit is designed to quantitatively measure formaldehyde present in tissue culture media and urine samples.
Brand DetectX®
Sample Type Urine, Tissue Culture Medium
Detection Method Fluorometric
Specificity Sample Types validated: Human Urine and Tissue Culture Media
Sensitivity 0.775 µM
Characteristics The Formaldehyde (HCHO) kit is designed to quantitatively measure formaldehyde present in buffer, tissue culture media and urine samples. A formaldehyde standard is provided to generate a standard curve for the assay. Standards or diluted samples are pipetted into a black microtiter plate. The fluorescent reaction is initiated with the DetectX Formaldehyde reagent. After a short incubation the emission of the generated fluorescent signal is detected in a microtiter plate reader capable of measuring 510nm fluorescence utilizing 450nm excitation wavelength. Formaldehyde is a toxic, colorless, flammable, strong-smelling gas. Materials containing formaldehyde can release formaldehyde gas or vapor into the air. Formaldehyde can also be released by burning wood, kerosene, natural gas, or cigarettes, from automobile emissions, and from natural processes. Studies have suggested that formaldehyde may affect the lymphatic and blood systems and that exposure to formaldehyde may cause health problems in humans. Industrial workers who help produce formaldehyde or formaldehyde-containing products, laboratory and health care professionals, and mortuary employees may be exposed to higher levels of formaldehyde than the general public. Exposure occurs primarily by inhaling formaldehyde gas or vapor from the air or by absorbing liquids containing formaldehyde through the skin. The National Cancer Institute has determined that there maybe an association between occupational exposure to formaldehyde and an increase in the risk of cancer. Several NCI studies have found that anatomists and embalmers, professions with potential exposure to formaldehyde, are at an increased risk for leukemia and brain cancer compared with the general population. For example, a multi-centered US study determined increased risk of nasopharyngeal cancer with formaldehyde exposure.
Components Black half Area 96 Well Plate 2 plates
Formaldehyde Standard 2,000 μM formaldehyde solution in special stabilizing solution. Outer container has formaldehyde absorbing pad. The standard is stable if kept tightly sealed. Keep Tightly Sealed 500 μL
DetectX® Formaldehyde reagent Special formulation of reagents to detect formaldehyde in solution. Contains 0.09% sodium azide as a preservative. 5 mL
Plate Sealers 2 each
Material not included A supply of distilled or deionized water free of formaldehyde.
Repeater pipet with disposable tips capable of dispensing 25 μL.
An incubator capable of accurately maintaining 37 °C.
Fluorescence 96 well plate reader capable of reading fluorescent emission at 510 nm, with excitation at 450 nm.
Set plate parameters for a 96-well Corning Costar 3694 plate. Software for converting raw relative fluorescent unit (FLU) readings from the plate reader and carrying out four parameter logistic curve (4PLC) fitting. it should be noted that most reactions should be compatible with the formaldehyde readout system. in systems where the amount of formaldehyde produced is low the amount of generated fluorescence will also be low. only plate readers that are capable of measuring dim fluorescent signals and having adjustable gain or filter settings may be compatible.
Background Formaldehyde (methanal), H C=O, is a colorless, flammable, strong-smelling gas. It is an important 2 industrial chemical used to manufacture building materials and to produce many household products. In the US approximately 3 x 109 kg are produced annually1. In addition, formaldehyde is commonly used as an industrial fungicide, germicide, and disinfectant, and as a preservative in mortuaries and medical laboratories. Materials containing formaldehyde can release formaldehyde gas or vapor into the air. Formaldehyde can also be released by burning wood, kerosene, natural gas, or cigarettes, from automobile emissions, and from natural processes. Formaldehyde can undergo rapid chemical changes immediately after absorption. Studies have suggested that formaldehyde may affect the lymphatic and blood systems and that exposure to formaldehyde may cause leukemia, particularly myeloid leukemia, in humans. Industrial workers who help to produce formaldehyde or formaldehyde-containing products, laboratory technicians, health care professionals, and mortuary employees may be exposed to higher levels of formaldehyde than the general public2. Exposure occurs primarily by inhaling formaldehyde gas or vapor from the air or by absorbing liquids containing formaldehyde through the skin. The National Cancer Institute (NCI) has determined that there is an association between occupational exposure to formaldehyde and an increase in the risk of cancer. Several NCI studies have found that anatomists and embalmers, professions with potential exposure to formaldehyde, are at an increased risk for leukemia and brain cancer compared with the general population. For example a multi-centered US study determined increased risk of nasopharyngeal cancer with formaldehyde exposure3
Application Notes Formaldehyde is identical across all species and cell types.
Urine and most types of tissue culture media (TCM) are compatible with this assay.
Assay Time 1 h
Plate 96 wells
Protocol A formaldehyde standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve.
Standards or diluted samples are pipetted into a black microtiter plate.
The fluorescent reaction is initiated with the DetectX® Formaldehyde reagent, which is pipetted into each well.
After a short incubation the emission of the generated fluorescent signal is detected in a microtiter plate reader capable of measuring 510 nm fluorescence utilizing 450 nm excitation wavelength.
The concentration of the formaldehyde in the sample is calculated, after making a suitable correction for the dilution of the sample, using software available with most fluorescence plate readers.
Reagent Preparation

Allow the kit reagents to come to room temperature for 30 minutes.
Ensure that all samples have reached room temperature and have been diluted as appropriate prior to running them in the kit.
Standard Preparation Label glass test tubes as #1 through #7.
Pipet 450 μL of water into tube #1 and 250 μL into tubes #2-#7.
Add 50 μL of the Formaldehyde stock solution to tube #1 and vortex completely.
Take 250 μL of the formaldehyde solution in tube #1 and add it to tube #2 and vortex completely.
Add 250 μL of tube #2 to tube #3 and vortex completely.
Repeat this serial dilution for tubes #4 through #7.
The concentration of formaldehyde in tubes 1 through 7 will be 200, 100, 50, 25, 12.5, 6.25 and 3.125 μM.
Water will be used as a sample blank.
Use all Standards within 2 hours of preparation.

Sample Preparation

Urine samples containing visible protein or particulates should be centrifuged or filtered prior to using. Urine samples must be diluted 1:4 with water by taking one part of sample and adding 3 parts of water prior to using in the kit. Any urine samples with formaldehyde concentrations outside the standard curve range should be diluted further with water to obtain readings within the standard curve. TCM samples should be diluted in TCM and read off a standard curve generated in the same TCM. Use all diluted samples within 2 hours of preparation.

Assay Procedure

We recommend that all standards and samples be run in duplicate to allow the end user to accurately determine formaldehyde concentrations.
1. A plate layout sheet has been included on the back page of the insert to aid proper sample and standard identification. Set plate parameters for a 96-well Corning Costar 3694 plate.
2. Pipet 50 μL of samples, water as the blank or standards into wells in the black plate.
3. Add 25 μL of the DetectX® Formaldehyde Reagent to each well using a repeater pipet.
4. Gently tap the sides of the plate to ensure adequate mixing of the reagents. Cover the plate with the plate sealer and press to seal adequately.
5. Incubate at 37 °C for 30 minutes. Room temperature incubation will yield approximately 75 % of the fluorescent signal generated with 37 °C incubation.
6. Read the fluorescent signal from each well in a plate reader capable of reading the fluorescent signal at 510 nm with excitation at 450 nm. Please contact your plate reader manufacturer for suitable filter sets.
7. Use the plate reader's built-in 4PLC software capabilities to calculate formaldehyde concentrations for each sample.

Calculation of Results

Average the duplicate FLU readings for each standard and sample.
Create a standard curve by reducing the data using the 4PLC fitting routine on the plate reader, after subtracting the mean FLUs for the zero standard.
The sample concentrations obtained should be multiplied by the dilution factor to obtain neat sample values.

Restrictions For Research Use only
Precaution of Use As with all such products, this kit should only be used by qualified personnel who have had laboratory safety instruction.
The complete insert should be read and understood before attempting to use the product.
Formaldehyde is a toxic, volatile, reactive chemical that can form adducts with proteins and nucleic acids.
It reacts with oxygen to form formic acid and so should be kept sealed and only used in well-ventilated laboratories.
For disposal, we suggest discarding all excess standards and samples in a 10% aqueous solution of sodium bisulfite, such as Sigma catalog number 13438.
Some of the components of this kit contain sodium azide as a preservative, which may react with lead or copper plumbing to form potentially explosive complexes.
When disposing of reagents always flush with large volumes of water to prevent azide build-up.
Storage 4 °C
Storage Comment All components of this kit should be stored at 4°C until the expiration date of the kit.
Supplier Images
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Product cited in: Pascual, Sotomayor, Rodríguez, Pérez-Köhler, Kühnhardt, Fernández-Gutiérrez, San Román, Bellón: "Cytotoxicity of Cyanoacrylate-Based Tissue Adhesives and Short-Term Preclinical In Vivo Biocompatibility in Abdominal Hernia Repair." in: PLoS ONE, Vol. 11, Issue 6, pp. e0157920, 2016 (PubMed).

McKenzie, Aruni, Johnson, Robles, Dou, Henry, Boskovic, Fletcher: "Metabolome variations in the Porphyromonas gingivalis vimA mutant during hydrogen peroxide-induced oxidative stress." in: Molecular oral microbiology, Vol. 30, Issue 2, pp. 111-27, 2015 (PubMed).