Maus Cholecystokinin ELISA Kit

Recommended Cholecystokinin Product (geliefert von: Anmelden zum Anzeigen )

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Cholecystokinin (CCK) ELISA Kits
  • CCK
  • cck-l
  • CCKN
  • cck-n
  • 2210420D18Rik
  • CARD3
  • CARDIAK
  • D4Bwg0615e
  • RICK
  • RIP2
  • cholecystokinin
  • cholecystokinin-L
  • cholecystokinin-N
  • receptor (TNFRSF)-interacting serine-threonine kinase 2
  • CCK
  • cck-l
  • cck-n
  • LOC100345534
  • cckn
  • Cck
  • LOC100355199
  • Ripk2
Reaktivität
Maus
Alternativen
Kits mit alternativen Reaktivitäten:
28
24
20
9
8
5
3
3
3
1
1
1
1
1
Methodentyp
Competition ELISA
Detektionsbereich
12.35-1000 pg/mL
Untere Nachweisgrenze
12.35 pg/mL
Applikation
ELISA
Optionen
Hersteller
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Bilder

Produktnummer ABIN415649
$ 545.68
Zzgl. Versandkosten $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Hersteller References Details
10.541325 ABIN367804 Sandwich ELISA Serum, Plasma, Tissue Homogenate 31.25-2000 pg/mL 31.25 pg/mL Anmelden zum Anzeigen
10.541325 ABIN424292 Competition ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 12.35-1000 pg/mL 12.35 pg/mL Anmelden zum Anzeigen
1 ABIN1979278 Competition ELISA Cell Culture Supernatant, Serum 0.1-1.000 pg/mL 0.1 pg/mL Anmelden zum Anzeigen 4
1 ABIN1979281 Competition ELISA Cell Culture Supernatant, Serum 0.1-1.000 pg/mL 0.1 pg/mL Anmelden zum Anzeigen 4
1 ABIN1979275 Competition ELISA Cell Culture Supernatant, Serum 0.1-1.000 pg/mL 0.1 pg/mL Anmelden zum Anzeigen 18
1 ABIN1568837 Competition ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.625-1000 pg/mL 15.625 pg/mL Anmelden zum Anzeigen
1 ABIN5652877 Competition ELISA Biological Fluids, Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate 12.35 pg/mL - 1000 pg/mL 12.35 pg/mL Anmelden zum Anzeigen
1 ABIN772912 Competition ELISA Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate, Body Fluids 5.0-100 ng/mL 5.0 ng/mL Anmelden zum Anzeigen
1 ABIN5522049 Sandwich ELISA Plasma, Serum, Biological Fluids, Tissue Homogenate 23.438-1500 pg/mL 23.438 pg/mL Anmelden zum Anzeigen
1 ABIN454698 Sandwich ELISA Serum, Tissue Homogenate, Biological Fluids Anmelden zum Anzeigen
1 ABIN955819 Sandwich ELISA 6.25-400 pg/mL 6.25 pg/mL Anmelden zum Anzeigen
1 ABIN2542892 Competition ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 12.35-1000 pg/mL 12.35 pg/mL Anmelden zum Anzeigen
1 ABIN4968013 Competition ELISA Cell Culture Supernatant, Plasma, Tissue Homogenate, Serum, Cell Lysate, Biological Fluids 15.625-1000 pg/mL 15.625 pg/mL Anmelden zum Anzeigen
1 ABIN2871373 Competition ELISA Anmelden zum Anzeigen
1 ABIN2871098 Competition ELISA Anmelden zum Anzeigen
1 ABIN4967877 Competition ELISA Cell Culture Supernatant, Plasma, Tissue Homogenate, Serum, Cell Lysate, Biological Fluids 15.625-1000 pg/mL 15.625 pg/mL Anmelden zum Anzeigen
1 ABIN4968011 Sandwich ELISA 23.438-1500 pg/mL 23.438 pg/mL Anmelden zum Anzeigen
1 ABIN2871376 Competition ELISA 0-120 ng/mL Anmelden zum Anzeigen
1 ABIN2871099 Competition ELISA Anmelden zum Anzeigen
1 ABIN5074233 Anmelden zum Anzeigen

General

Antigen Cholecystokinin (CCK) ELISA Kits
Reaktivität Maus
Kits mit alternativen Reaktivitäten:
(28), (24), (20), (9), (8), (5), (3), (3), (3), (1), (1), (1), (1), (1)
Methodentyp Competition ELISA
Detektionsbereich 12.35-1000 pg/mL
Untere Nachweisgrenze 12.35 pg/mL
Applikation ELISA
Hersteller Anmelden zum Anzeigen

Produktdetails Cholecystokinin ELISA Kit

Antigendetails Anwendungsinformationen Handhabung Bilder
Verwendungszweck The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of CCK in Serum,Plasma,Tissue Homogenate,Cell Lysate,Cell Culture Supernatant,Biological Fluids
Proben Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids
Nachweismethode Colorimetric
Analytische Methode Quantitative
Spezifität

This assay has high sensitivity and excellent specificity for detection of Cholecystokinin (CCK).
No significant cross-reactivity or interference between Cholecystokinin (CCK) and analogues was observed.

Kreuzreaktivität (Details) No significant cross-reactivity or interference between Cholecystokinin (CCK) and analogues was observed.
Sensitivität 4.82 pg/mL
Bestandteile
  • Pre-coated, ready to use 96-well strip plate
  • Plate sealer for 96 wells
  • Standard Diluent
  • Assay Diluent A
  • Assay Diluent B
  • Stop Solution
  • Standard
  • Detection Reagent A
  • Detection Reagent B
  • TMB Substrate
  • Wash Buffer (30 × concentrate)
  • Instruction manual
Benötigtes Material
  • Microplate reader with 450 nm filter.
  • Precision single or multi-channel pipettes and disposable tips.
  • Eppendorf Tubes for diluting samples.
  • Deionized or distilled water.
  • Absorbent paper for blotting the microtiter plate.
  • Container for Wash Solution

Antigendetails

Produktdetails Cholecystokinin ELISA Kit Anwendungsinformationen Handhabung Bilder zurück nach oben
Antigen
Andere Bezeichnung CCK (CCK ELISA Kit Abstract)
UniProt P09240
Pathways T-Zell Rezeptor Signalweg, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, Positive Regulation of Endopeptidase Activity, Toll-Like Receptors Cascades, Feeding Behaviour

Anwendungsinformationen

Produktdetails Cholecystokinin ELISA Kit Antigendetails Handhabung Bilder zurück nach oben
Applikationshinweise
  • Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
  • The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
  • Kits from different batches may be a little different in detection range, sensitivity and color developing time.
  • Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
  • Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
  • There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
  • Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
  • Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
  • Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
  • Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.
Kommentare

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Probenmenge 50 μL
Testdauer 2 h
Plattentyp Pre-coated
Protokoll This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Cholecystokinin (CCK) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Cholecystokinin (CCK) and unlabeled Cholecystokinin (CCK) (Standards or samples) with the pre-coated antibody specific to Cholecystokinin (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 Cholecystokinin (CCK) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Cholecystokinin (CCK) in the sample.
Aufbereitung der Reagenzien
  • Bring all kit components and samples to room temperature (18-25°C) before use.
  • Standard - Reconstitute the Standard with 1.0mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 1,000pg/mL. Please prepare 5 tubes containing 0.6mL Standard Diluent and produce a triple dilution series. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 1,000pg/mL, 333.33pg/mL, 111.11pg/mL, 37.04pg/mL, 12.35pg/mL, and the last EP tubes with Standard Diluent is the blank as 0pg/mL.
  • Assay Diluent A and Assay Diluent B - Dilute 6mL of Assay Diluent A or B Concentrate(2×) with 6mL of deionized or distilled water to prepare 12 mL of Assay Diluent A or B. (In fact, more than 6mL Assay Diluent A and Assay Diluent B are contained in the bottles. Therefore, in every test, please precisely pipette required amount of Diluent and make double dilution in a new container. The prepared working dilution cannot be frozen.)
  • Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with working Assay Diluent A or B, respectively (1:100).
  • Wash Solution - Dilute 20mL of Wash Solution concentrate (30×) with 580mL of deionized or distilled water to prepare 600 mL of Wash Solution (1×).
  • TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.
Note:
  • Making serial dilution in the wells directly is not permitted.
  • Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37°C directly.
  • Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
  • Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10µL for once pipetting.
  • The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
  • If crystals have formed in the Wash Solution concentrate (30×), warm to room temperature and mix gently until the crystals are completely dissolved.
  • Contaminated water or container for reagent preparation will influence the detection result.
Probennahme 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 Lysate: 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.

Cell Culture Supernatant: 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.

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.
Aufbereitung der Proben
  • We are only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
  • Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments. Sample should be diluted by 0.01mol/L PBS(PH=7.0-7.2).
  • If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
  • Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts from certain chemicals.
  • Due to the possibility of mismatching between antigen from other origin and antibody used in our kits (e.g.antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
  • Influenced by the factors including cell viability, cell number or sampling time, samples from cell culture supernatant may not be detected by the kit.
  • Fresh samples without long time storage is recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.
Testdurchführung
  1. Prepare all reagents, samples and standards,
  2. Add 50μL standard or sample to each well.
    And then add 50μL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37 °C,
  3. Aspirate and wash 3 times,
  4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
  5. Aspirate and wash 5 times,
  6. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
  7. Add 50μL Stop Solution. Read at 450 nm immediately.
Ergebnisberechnung 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.
Testpräzision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cholecystokinin (CCK) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cholecystokinin (CCK) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Beschränkungen Nur für Forschungszwecke einsetzbar

Handhabung

Produktdetails Cholecystokinin ELISA Kit Antigendetails Anwendungsinformationen Bilder zurück nach oben
Vorsichtsmaßnahmen The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
Handhabung

The stability of 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.
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.

Lagerung 4 °C
Informationen zur Lagerung
  • For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
  • For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
    Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
  • 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.
Haltbarkeit 6 months

Bilder

Produktdetails Cholecystokinin ELISA Kit Antigendetails Anwendungsinformationen Handhabung zurück nach oben
Bilder des Herstellers
ELISA image for Cholecystokinin (CCK) ELISA Kit (ABIN415649) Cholecystokinin (CCK) ELISA Kit