|+1 404 474 4654|
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Cholecystokinin (CCK) ELISA Kit
|Synonyms||CCK-CHR, CCK, MGC140193, cck-l, CCKN, cck-n, MGC117187|
|Certificates||ISO 9001:2008, ISO 13485:2003|
|Price||729.47 $ Plus shipping costs $45.00|
|Availability||Will be delivered in 9 to 11 Business Days|
|Method type||Competition ELISA|
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between human CCK and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross- reactivity detection between human CCK and all the analogues, therefore, cross reaction may still exist.|
|Sample Type||Serum, Plasma, Biological Fluids, Tissue Homogenate, Cell Culture Supernatant|
|Plate||Pre-coated 12 × 8 strip|
|Specificity||This assay has high sensitivity and excellent specificity for detection of human CCK.|
|Sensitivity||The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by subtracting two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.|
|Minimum Detection Limit||4.55 pg/mL|
|Detection Range||12.35-1,000 pg/mL|
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level human CCK were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level human CCK were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
The standard curve concentrations used for the ELISA’s were 1,000pg/mL, 333.33pg/mL, 111.11pg/mL, 37.04pg/mL, 12.35pg/mL.The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 3 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C).
|Principle||The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of CCK in human serum, plasma, tissue homogenates, cell culture supernates and other biological fluids.|
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for human CCK has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled human CCK and unlabeled human CCK (Standards or samples) with the pre-coated antibody specific for human CCK. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of CCK in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of CCK in the sample.
Not all sample types may be valid for this kit. Please look up which sample types apply.
Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Tissue homogenates: The preparation of tissue homogenates will vary depending upon tissue type. For this assay, rinse tissues in ice-cold PBS (0.02mol/L,pH 7.0-7.2) to remove excess blood thoroughly and weigh before homogenization. Mince the tissues to small pieces and homogenize them in 5-10 mL of PBS with a glass homogenizer on ice (Micro Tissue Grinders work, too). Sonicate the resulting suspension with an ultrasonic cell disrupter or subject it to two freeze-thaw cycles to further break the cell membranes. Centrifugate the homogenates for 5 minutes at 5000 × g. Remove the supernate and assay immediately or aliquot and store at -20°C.
Cell lysates: Cells must be lysed before assaying according to the following directions. Adherent cells should be detached with trypsin and then collected by centrifugation (suspension cells can be collected by centrifugation directly). Wash cells three times in cold PBS. Resuspend cells in PBS (1×) and subject them to ultrasonication for 4 times (or Freeze cells at -20 °C. Thaw cells with gentle mixing. Repeat the freeze/thaw cycle for 3 times.) Centrifuge at 1500 × g for 10 minutes at 2 - 8°C to remove cellular debris.
Platelet-poor plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g at 2-8°C within 30 minutes of collection. It is recommended to centrifuge samples for 10 minutes at 10,000 × g for complete platelet removal. Remove plasma and assay (see activation procedure) immediately or store samples in aliquot at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.
Saliva: Collect sample in centrifuge tube. To clarify, freeze sample at -70°C for 1 hour. Thaw sample on ice, and centrifuge at 2,000 × g for 10 minutes. Transfer clarified supernatant to clean tube for use in the assay. Alternatively, the clarified saliva samples can be frozen at -20°C and assayed at a later date. It is recommended that the saliva sample be aliquoted to convenient volumes prior to storing at -20°C to avoid multiple freeze thaw cycles.
Seminal plasma: Collect semen samples using a sterile container, then allow it to liquefy at room temperature or 37°C. After liquefaction, centrifuge at 4000 rpm for 10 min. Remove the supernate and assay immediately or store samples at -20°C.
Sperm: Collect semen samples using a sterile container, then allow it to liquefy at room temperature or 37°C. After liquefaction, centrifuge at 2,000 × g for 10-15 minutes and remove seminal plasma, wash precipitated protein and centrifuge for three times. Subject the precipitated protein to ultrasonication, centrifuge at 2,000 × g for 10-15 minutes. Remove the supernate and assay immediately or store samples at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.
Urine: Aseptically collect the first urine of the day (mid-stream) , voided directly into a sterile container. Centrifuge to remove particulate matter, assay immediately or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles.
Milk, cell culture supernates and other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.Tears: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.
Lung lavage fluid: Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
Cerebrospinal fluid: Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze-thaw cycles.
Biological agents: Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze-thaw cycles.
Sweat: Collect sweat using a collection device or equivalent. Assay immediately or aliquot and store samples at -20°C. Avoid repeated freeze-thaw cycles.
|Calculation of Results||
This assay employs the competitive inhibition enzyme immunoassay technique, so there is an inverse correlation between CCK concentration in the sample and the assay signal intensity. Average the duplicate readings for each standard, control, and samples. Create a standard curve on log-log or semi-log graph paper, with the log of CCK concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the standard points and it can be determined by regression analysis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the log of concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. The O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects). Typical standard curve below is provided for reference only.
|Handling Advice||To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.|
|Precaution of Use||The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.|
|Material not included||
|Expiry Date||The expiry date is stated on the label.|
|Restrictions||For Research Use only|