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#DICL-250 QuantiChrom™ Chloride Assay Kit
Product name : QuantiChrom™ Chloride Assay Kit
Catalog number : DICL-250
Supplier name : Bioassays
Data sheet: Ask more or other datasheet now !
About this Product :QuantiChrom™ Chloride Assay Kit antibody storage GENTAUR recommends for long therm storage to freeze at -24 C. For short time storage up to 30 days we suggest fridge storage at 1 to 10 C. Prevent multiple freeze taw cycles of QuantiChrom™ Chloride Assay Kit.
More Details about
For quantitative determination of chloride ion and evaluation of drug effects on chloride metabolism.
Samples: biological, food and environment.
Procedure: 5 min.
Size: 250 tests.
Detection limit: 0.7 mg/dL (0.2 mM).
Chloride is the major extracellular anion in human body fluids. Chloride
plays a key role in maintaining proper water distribution, osmotic
pressure and electrolyte balance. Low chloride concentrations may be
found with prolonged vomiting, extensive burns, metabolic acidosis,
Addisonia crisis and renal diseases. Elevated chloride concentrations are
associated with dehydration, congestive heart failure, hyperventilation
and urinary obstructions. Determination of chloride in sweat is useful in
diagnosing cystic fibrosis.
Simple, direct and automation-ready procedures for measuring chloride
concentration in biological samples are becoming popular in Research
and Drug Discovery. BioAssay Systems' chloride assay kit is designed to
measure chloride directly in biological samples without any pretreatment.
The improved Fried method utilizes mercuric 2,4,6-tripyridyl-s-triazine,
which forms a colored complex specifically with chloride. The intensity of
the color, measured at 610nm, is directly proportional to the chloride
concentration in the sample. The optimized formulation substantially
reduces interference by substances in the raw samples.
Sensitive and accurate. Use as little as 5 μL samples. Linear detection
range 0.7 mg/dL (0.2mM) to 35 mg/dL (10mM) Cl- in 96-well plate assay.
Simple and high-throughput. The procedure involves addition of a
single working reagent and incubation for 5 min. Can be readily automated
as a high-throughput assay for thousands of samples per day.
Improved reagent stability and versatility. The optimized formulation has
greatly enhanced reagent and signal stability. Cuvet or 96-well plate assay.
Low interference in biological samples. No pretreatments are needed.
Assays can be directly performed on raw biological samples i.e., in the
presence of lipid, protein and minerals such as magnesium, iron and zinc.
Direct Assays: Cl- in serum, plasma, urine, saliva, sweat etc.
Drug Discovery/Pharmacology: effects of drugs on chloride metabolism.
Food and Beverages: chloride determination.
Environment: chloride determination in water and soil.
KIT CONTENTS (250 tests in 96-well plates)
Reagent: 50 mL Chloride standard: 1 mL 35 mg/dL Cl-
Storage conditions. The kit is shipped at room temperature. Store
Reagent and Standard at 4°C. Shelf life: 12 months after receipt.
Precautions: reagents are for research use only. Normal precautions for
laboratory reagents should be exercised while using the reagents.
MATERIALS REQUIRED, BUT NOT PROVIDED
Pipeting devices and accessories (e.g. 5 μL), clear flat-bottom 96-
well plates and plate reader, or spectrophotometer and cuvets.
Sample Type: serum
References: Borenshtein, D et al (2009). Decreased expression of colonic Slc26a3 and carbonic anhydrase iv as a cause of fatal infectious diarrhea in mice. Infect Immun. 77(9):3639-50
Pubmed ID: 19546193
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=19546193
Abstract: Citrobacter rodentium causes epithelial hyperplasia and colitis and is used as a model for enteropathogenic and enterohemorrhagic Escherichia coli infections. Little or no mortality develops in most inbred strains of mice, but C3H and FVB/N mice exhibit fatal outcomes of infection. Here we test the hypothesis that decreased intestinal transport activity during C. rodentium infection results in fatality in C3H/HeOu and FVB/N mice. Susceptible strains were compared to resistant C57BL/6 mice and to inbred strains SWR and SJL of Swiss origin, which have not been previously characterized for outcomes of C. rodentium infection. Mortality in susceptible strains C3H/HeOu and FVB/N was associated with significant fluid loss in feces, a remarkable downregulation of Slc26a3 and carbonic anhydrase IV (CAIV) message and protein expression, retention of chloride in stool, and hypochloremia, suggesting defects in intestinal chloride absorption. SWR, SJL, and C57BL/6 mice were resistant and survived the infection. Fluid therapy fully prevented mortality in C3H/HeOu and FVB/N mice without affecting clinical disease. Common pathogenic mechanisms, such as decreased levels of expression of Slc26a3 and CAIV, affect intestinal ion transport in C. rodentium-infected FVB and C3H mice, resulting in profound electrolyte loss, dehydration, and mortality. Intestinal chloride absorption pathways are likely a potential target for the treatment of infectious diarrhea.
[PubMed - indexed for MEDLINE] PMCID: PMC2738044
Sample Type: junction liquid
References: Peng, S et al (2010). Minimal effects of VEGF and anti-VEGF drugs on the permeability or selectivity of RPE tight junctions. Invest Ophthalmol Vis Sci. 51(6):3216-25
Pubmed ID: 20042644
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=20042644
PURPOSE: Bevacizumab and ranibizumab are currently used to treat age-related macular degeneration by neutralizing vascular endothelial growth factor (VEGF). In this study, the potential side effects on the outer blood-retinal barrier were examined.
METHODS: Human fetal RPE (hfRPE) cells were used because they are highly differentiated in culture. The claudin composition of RPE tight junctions was determined by RT-PCR, immunoblot analysis, and immunofluorescence. ELISA assays monitored the secretion and trafficking of VEGF and a fluid-phase marker, methylpolyethylene glycol (mPEG). Tight junction functions were assessed by the conductance of K(+) and Na(+) (derived from the transepithelial electrical resistance, TER) and the flux of NaCl and mPEG.
RESULTS: Claudin-3, claudin-10, and claudin-19 were detected in RPE tight junctions. VEGF was secreted in equal amounts across the apical and basolateral membranes, but the apical membrane was more active in endocytosing and degrading VEGF. Exogenous VEGF and mPEG crossed the RPE monolayer by transcytosis, predominantly in the apical-to-basal direction. RPE tight junctions were selective for K(+), but did not discriminate between Na(+) and Cl(-). VEGF, bevacizumab, and ranibizumab had minimal effects on TER, permeation of mPEG, and selectivity for K(+), Na(+), and Cl(-). They had minimal effects on the expression and distribution of the claudins.
CONCLUSIONS: RPE has mechanisms for maintaining low concentrations of VEGF in the subretinal space that include endocytosis and degradation and fluid-phase transcytosis in the apical-to-basal direction. RPE tight junctions are selective for K(+) over Na(+) and Cl(-). Permeability and selectivity of the junctions are not affected by VEGF, bevacizumab, or ranibizumab.
[PubMed - indexed for MEDLINE] PMCID: PMC2891474
Sample Type: sulfite oxidase
References: Rajapakshe, A et al (2010). Characterization of chloride-depleted human sulfite oxidase by electron paramagnetic resonance spectroscopy: experimental evidence for the role of anions in product release. Biochemistry 49(25):5154-9
Pubmed ID: 20491442
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=20491442
Abstract: The Mo(V) state of the molybdoenzyme sulfite oxidase (SO) is paramagnetic and can be studied by electron paramagnetic resonance (EPR) spectroscopy. Vertebrate SO at pH <7 and >9 exhibits characteristic EPR spectra that correspond to two structurally different forms of the Mo(V) active center termed the low-pH (lpH) and high-pH (hpH) forms, respectively. Both EPR forms have an exchangeable equatorial OH ligand, but its orientation in the two forms is different. It has been hypothesized that the formation of the lpH species is dependent on the presence of chloride. In this work, we have prepared and purified samples of the wild type and various mutants of human SO that are depleted of chloride. These samples do not exhibit the typical lpH EPR spectrum at low pH but rather exhibit spectra that are characteristic of the blocked species that contains an exchangeable equatorial sulfate ligand. Addition of chloride to these samples results in the disappearance of the blocked species and the formation of the lpH species. Similarly, if chloride is added before sulfite, the lpH species is formed instead of the blocked one. Qualitatively similar results were observed for samples of sulfite-oxidizing enzymes from other organisms that were previously reported to form a blocked species at low pH. However, the depletion of chloride has no effect upon the formation of the hpH species.
[PubMed - indexed for MEDLINE]PMCID: PMC2890295
Sample Type: urine
References: Grünberg, W et al (2011). Periparturient effects of feeding a low dietary cation-anion difference diet on acid-base, calcium, and phosphorus homeostasis and on intravenous glucose tolerance test in high-producing dairy cows.
Pubmed ID: 21257041
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=21257041
Abstract: Feeding rations with low dietary cation-anion difference (DCAD) to dairy cows during late gestation is a common strategy to prevent periparturient hypocalcemia. Although the efficacy of low-DCAD rations in reducing the incidence of clinical hypocalcemia is well documented, potentially deleterious effects have not been explored in detail. The objective of the study presented here was to determine the effect of fully compensated metabolic acidosis on calcium and phosphorus homeostasis, insulin responsiveness, and insulin sensitivity as well as on protein metabolism. Twenty multiparous Holstein-Friesian dairy cows were assigned to 1 of 2 treatment groups and fed a low-DCAD ration (DCAD = -9 mEq/100g, group L) or a control ration (DCAD = +11 mEq/100g, group C) for the last 3 wk before the expected calving date. Blood and urine samples were obtained periodically between 14 d before to 14 d after calving. Intravenous glucose tolerance tests and 24-h volumetric urine collection were conducted before calving as well as 7 and 14 d postpartum. Cows fed the low-DCAD ration had lower urine pH and higher net acid excretion, but unchanged blood pH and bicarbonate concentration before calving. Protein-corrected plasma Ca concentration 1 d postpartum was higher in cows on the low-DCAD diet when compared with control animals. Urinary Ca and P excretion was positively associated with urine net acid excretion and negatively associated with urine pH. Whereas metabolic acidosis resulted in a 6-fold increase in urinary Ca excretion, the effect on renal P excretion was negligible. A more pronounced decline of plasma protein and globulin concentration in the periparturient period was observed in cows on the low-DCAD diets resulting in significantly lower total protein and globulin concentrations after calving in cows on low-DCAD diets. Intravenous glucose tolerance tests conducted before and after calving did not reveal group differences in insulin response or insulin sensitivity. Our results indicate that fully compensated metabolic acidosis increased the Ca flux resulting in increased urinary calcium excretion before calving and increased plasma Ca concentration on the day after calving, whereas the effect on P homeostasis was unlikely to be clinically relevant. The clinical relevance of the effect of metabolic acidosis on the plasma protein and globulin concentration is unclear but warrants further investigation.
Copyright Â© 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
[PubMed - indexed for MEDLINE]
Sample Type: serum
References: Jensen, SB et al (2008). Adjuvant chemotherapy in breast cancer patients induces temporary salivary gland hypofunction. Oral Oncol. 44(2):162-73
Pubmed ID: 17588802
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=17588802
Abstract: It is an open question if chemotherapy (CT) per se imposes adverse effects on salivary gland function. The aim of the present study was to investigate effects of CT on salivary function in breast cancer patients during and after adjuvant CT. Forty-five breast cancer patients, eligible for adjuvant CT with CEF or CMF (cyclophosphamide, epirubicin or methotrexate, 5-fluorouracil) were followed before, during, six months and one year after CT. Findings were compared to those in a control group of 31 breast cancer patients not receiving CT. Flow rates and compositions of unstimulated and stimulated whole saliva as well as stimulated parotid saliva (UWS, SWS and SPS) were measured. Feeling of oral dryness and saliva-related complaints were registered. UWS and SWS flow rates decreased during CT (p<0.001 and p<0.01). UWS remained lower six months after, but reached baseline level within one year. SPS flow rate was not significantly affected, suggesting that the decrease in whole saliva production is accounted for by decreased acinar saliva formation by the submandibular glands. Twenty patients (44%) suffered from hyposalivation (UWS < or = 0.1 ml/min and/or SWS < or = 0.5 ml/min) during CT. Xerostomia scores rose during CT and stayed elevated one year after treatment. CT also induced compositional changes by slightly increasing salivary sodium and chloride concentrations as well as decreasing inorganic phosphate concentrations in spite of lower or unchanged flow rates, implying that ductal modification mechanisms are affected. UWS and SWS total protein output and UWS secretory IgA output decreased in response to CT. Thus, the results suggest that acinar and ductal cell functions are affected by adjuvant CT. These adverse drug reactions are temporary, as salivary findings generally returned to baseline values within one year following treatment.
[PubMed - indexed for MEDLINE]
Sample Type: blood
Species: broiler chick
References: Ahmad, T et al (2009). Influence of varying dietary electrolyte balance on broiler performance under tropical summer conditions. J Anim Physiol Anim Nutr (Berl). 93(5):613-21
Pubmed ID: 19141104
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=19141104
Abstract: A 6-week feeding trial was conducted to determine the effects of varying dietary electrolyte balance (DEB) on growth performance and physiological responses in broiler chicks reared during hot summer months (26.1-37.5 degrees C). The average minimum and maximum room temperatures recorded from 14 to 42 days of age were 26.1 and 37.5 degrees C, respectively, with relative humidity ranging from 51% to 55%. Five DEB treatments supplying 0, 50, 150, 250 and 350 (Na(+)+K(+)-Cl(-)) mEq/kg of diet were prepared by adding NaHCO(3) and/or NH(4)Cl to basal diet that contained 185 mEq/kg from 1 to 28 days and 172 mEq/kg from 29 to 42 days. These DEB diets were randomly offered to six experimental units of 20 chicks each. Improved 42-day body weight gain (BWG), feed:gain (F:G) and reduced mortality was noted with DEB 50, 150 and 250 mEq/kg compared to DEB 0 and 350 mEq/kg. A quadratic effect of increasing DEB level was observed on BWG and F:G, whereas the effect was linear for feed intake, water intake and litter moisture. The DEB 50, 150 and 250 maintained better blood pH, pCO(2) and HCO(3) concentrations during acute heat stress periods than that of 0 and 350. The blood heterophil and lymphocyte percent as well as heterophil:lymphocyte were remained unaffected. The lowest DEB (0 mEq/kg) diet reduced the feed intake and promoted metabolic acidosis. At 350 mEq/kg DEB, water intake and metabolic alkalosis were promoted probably due to high levels of dietary NaHCO(3). The study concluded that overall better performance was recorded with DEB 50, 150 and 250 mEq/kg. These results indicated that single optimal DEB value could not be recommended to combat heat stress in broilers.
[PubMed - indexed for MEDLINE]
Sample Type: plasma
References: Everaert, N et al (2011). Regulatory capacities of a broiler and layer strain exposed to high CO2 levels during the second half of incubation. Comp Biochem Physiol A Mol Integr Physiol. 158(2):215-20
Pubmed ID: 21056110
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=21056110
Abstract: It has been shown that during embryonic chicken (Gallus gallus) development, the metabolism of broiler embryos differs from that of layers in terms of embryonic growth, pCO2/pO2 blood levels, heat production, and heart rate. Therefore, these strains might adapt differently on extreme environmental factors such as exposure to high CO2. The aim of this study was to compare broiler and layer embryos in their adaptation to 4% CO2 from embryonic days (ED) 12 to 18. Due to hypercapnia, blood pCO2 increased in both strains. Blood bicarbonate concentration was ~10 mmol/L higher in embryos exposed to high CO2 of both strains, while the bicarbonates of broilers had ~5 mmol/L higher values than layer embryos. In addition, the pH increased when embryos of both strains were exposed to CO2. Moreover, under CO2 conditions, the blood potassium concentration increased in both strains significantly, reaching a plateau at ED14. At ED12, the layer strain had a higher increase in CAII protein in red blood cells due to incubation under high CO2 compared to the broiler strain, whereas at ED14, the broiler strain had the highest increase. In conclusion, the most striking observation was the similar mechanism of broiler and layer embryos to cope with high CO2 levels. Copyright Â© 2010 Elsevier Inc. All rights reserved. PM [PubMed - indexed for MEDLINE]
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