Sample
Serum, urine or saliva.
Intended Use
The Cotinine ELISA Kit is intended for the measurement of Cotinine in serum, urine or saliva.
Contents of Kit
1. Microwell coated with polyclonal Ab to Cotinine. 12 × 8
2. Standard Set (ready to use). 0.5 mL
3. Cotinine HRP Enzyme Conjugate (ready to use). 12 mL
4. TMB Substrate (ready to use). 12 mL
5. Stop Solution (ready to use). 12 mL
6. Wash Concentrate. 20×, 1 bottle, 25 mL
Storage
1. Store the kit at 2 - 8°C.
2. Keep microwells sealed in a dry bag with desiccants.
3. The reagents are stable until expiration of the kit.
4. Do not expose test reagents to heat, sun, or strong light.
Performance Characteristics
Accuracy20 urine samples from non-smokers were screened with this Cotinine ELISA method. All 20 samples screened negative with the ELISA method. 15 samples from smokers which contained various amounts of Cotinine were screened with This Cotinine Direct ELISA Kit. All 15 samples showed a presence of Cotinine at a level greater than 500 ng/mL. Three urine samples submitted by individuals exposed to passive inhalation for over 30 days all showed levels of 5 to 10 ng/mL of Cotinine when extrapolated of a dose response curve.
SensitivityAssay sensitivity based on the minimum Cotinine concentration required to produce a three-standard deviation from assay Ao is 1 ng/mL.
SpecificityThe specificity of this Cotinine ELISA was determined by generating inhibition curves for each of the compounds listed below the antisera cross-reactivity below.

Detection Range
5-100 ng/mL
General Description
Exposure to tobacco smoke can be detected by measuring nicotine and its metabolites. Nicotine has a short half life and is not used as a marker for tobacco smoke exposure. Cotinine due to its longer half life has been used in research as a reliable marker for smoking status and smoking cessation studies. The Cotinine Direct ELISA Kit is designed for the detection Cotinine in serum and urine. It can also be adapted for other fluids.
Citations
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Ameliorative Effect of Gum Acacia on Hookah Smoke-Induced Testicular Impairment in Mice
Ali BH, Al-Salam S, Balushi KAA, Za'abi MA, Adham SA, Beegam S, Yuvaraju P, Manoj P, Nemmar A
Applications: ELISA
Reactive species: Human
"Abstract: We investigated some reproductive actions of hookah smoke (HS) exposure (30 min/day, for 30 days) in male mice, and the possible mitigative effect of the prebiotic agent gum acacia (GA) thereon. Control mice were air-exposed (AE). Twenty-four hours after the last exposure, the levels of some plasma reproductive hormones, biochemical markers of inflammation, oxidative and nitrosative stress and testicular histopathology were assessed. The urinary level of cotinine, a major nicotine metabolite, was also measured. HS exposure induced significant decreases in testosterone, estradiol, luteinizing hormone, and androgen binding protein, as well as glutathione reductase activity and levels of nitrite and total nitrite. Plasma inhibin B, alkaline phosphatase, lipopolysaccharide binding protein, uric acid, lactate dehydrogenase, lipid peroxidation, 8-oxo-2'-deoxyguanosine, and cytochrome C were significantly increased following HS exposure. In testicular homogenate, nuclear factor-κB (NF-ĸB), nuclear factor erythroid 2-related factor 2 (Nrf2), interleukin- 6 (IL-6), interleukin-1β (IL-1β), transforming growth factor-β1(TGF- β1), and tumor necrosis factor-α (TNF- α) were all significantly elevated, and the steroidogenic acute regulatory protein (StAR) significantly decreased. Histopathologically, there was slight impairment and disorganization of spermatogenesis. Urinary cotinine concentration was elevated significantly in the HS-exposed group compared with the air-exposed group. GA co-administration mitigated the adverse actions of HS measured. In conclusion, daily exposure to HS at the above dose induced adverse actions on the reproductive system of male mice. GA co-administration significantly mitigated these effects by reducing the inflammation, oxidative and nitrosative stress, via a mechanism involving Nrf2, and reduction of StAR expression."
Article snippet: The urinary level of cotinine was assayed by an ELISA kit from Creative Diagnostics (Shirley, NY, USA).
Figure 1. Urine cotinine level of mice exposed to air (AE) or hookah smoke (HS) with or without GA treatment.
Chronic Water-Pipe Smoke Exposure Induces Injurious Effects to Reproductive System in Male Mice
Badreldin H Ali, Khalid A Al Balushi, Mohammed Ashique, Asem Shalaby, Mohammed A Al Kindi, Sirin A Adham, Turan Karaca, Sumaya Beegam, Priya Yuvaraju, Abderrahim Nemmar
Applications: ELISA
Reactive species: Human
"Abstract: There is a global increase in the popularity of water-pipe tobacco smoking including in Europe and North America. Nevertheless, little is known about the male reproductive effects of water-pipe smoke (WPS), especially after long-term exposure. Here, we assessed effects of WPS exposure (30 min/day) in male mice for 6 months. Control mice were exposed to air-only for the same period of time. Twenty-four hours after the last exposure, testicular histopathology, and markers of inflammation and oxidative stress, and the tyrosine–protein kinase vascular endothelial growth factor receptor 1 (VEGFR1) were assessed in testicular homogenates. Moreover, plasma testosterone, estradiol, and luteinizing hormone (LH) concentrations were also measured. Chronic WPS exposure induced a significant decrease of testosterone and estradiol, and a slight but significant increase of LH. Glutathione reductase, catalase, and ascorbic acid were significantly decreased following WPS exposure. Plasma concentration of leptin was significantly decreased by WPS exposure, whereas that of tumor necrosis factor α and interleukin 6 was significantly increased. Histopathological analysis of the testes revealed the presence of a marked reduction in the diameter of the seminiferous tubules with reduced spermatogenesis. Transmission electron microscopy examination showed irregular thickening and wrinkling of the basement membranes with abnormal shapes and structures of the spermatozoa. VEGFR1 was overexpressed in the testis of the mice exposed to WPS and was not detected in the control. The urine concentration of cotinine, the predominant metabolite of nicotine, was significantly increased in the WPS-exposed group compared with the control group."
Article snippet: The concentrations of cotinine in urine (ELISA kit from Creative Diagnostics, Shirley, NY, USA) were measured after the last exposure to either WPS or air on urine samples collected overnight using metabolic cages for urine collection.
Figure 1. The concentrations of cotinine in plasma (A) and urine (B) following air exposure (AE) of control mice (open columns), and mice exposed to water-pipe smoke (WPS) for 30 min per day for 6 months (filled columns).
Waterpipe Tobacco Smoke Inhalation Triggers Thrombogenicity, Cardiac Inflammation and Oxidative Stress in Mice: Effects of Flavouring
Abderrahim Nemmar, Suhail Al-Salam, Sumaya Beegam, Priya Yuvaraju, Nur Elena Zaaba, Javed Yasin, Badreldin H Ali
Applications: ELISA
Reactive species: Human
"Abstract: The consumption of water-pipe smoking (WPS) has been promoted by the use of flavoured tobacco. However, little is known about the impact of flavouring on the cardiovascular toxicity induced by WPS inhalation. Here, we compared the cardiovascular effects and underlying mechanism of actions of plain (P) (unflavoured) versus apple-flavoured (AF) WPS (30 minutes/day, 5 days/week for 1 month) in mice. Control mice were exposed to air. Both P- and AF-WPS inhalation induced an increase in systolic blood pressure, thrombogenicity and plasma concentration of fibrinogen and von Willebrand factor. In heart homogenates, AF-WPS inhalation caused an increase of 8-isoprostane and a decrease in the levels of reduced glutathione (GSH) and superoxide dismutase (SOD). Nevertheless, P-WPS decreased only the activity of SOD. The concentrations of tumour necrosis factor α and interleukin 1β were increased only in heart homogenates of mice exposed to AF-WPS."
Article snippet: The quantification of urinary cotinine concentrations was performed using an ELISA kit purchased from Creative Diagnostics (Shirley, NY, USA).
Figure 1. Effects of plain water-pipe smoke (P-WPS) or apple-flavoured (AF)-WPS on cotinine concentrations in urine.