Vasopressin ELISA Kit (Arginine Vasopressin)

Details for Product AVP ELISA Kit No. ABIN2506989, Anbieter: Anmelden zum Anzeigen
Antigen
  • ADH
  • ARVP
  • AVP-NPII
  • AVRP
  • VP
  • Vp
  • Vsp
  • DI
  • Vas
  • adh
  • arvp
  • avrp
  • avp-npii
  • copeptin
  • vasotocin
  • arginine vasopressin
  • AVP
  • Avp
  • avp
Reaktivität
Rind (Kuh)
Alternativen
Kits mit alternativen Reaktivitäten:
33
30
24
7
6
4
4
4
1
1
1
1
1
1
Methodentyp
Competition ELISA
Detektionsbereich
2.4-10000 pg/mL
Untere Nachweisgrenze
2.4 pg/mL
Applikation
ELISA
Optionen
Hersteller
Anmelden zum Anzeigen
Hersteller Produkt- Nr.
Anmelden zum Anzeigen
Verwendungszweck The kit is a competitive enzyme immunoassay for the in vitro quantitative measurement of Vasopressin in bovine serum, plasma, and other biological fluids not being tested
Marke EasyTest™
Proben Serum, Plasma
Analytische Methode Quantitative
Nachweismethode Colorimetric
Spezifität This assay has high sensitivity and excellent specificity for detection of bovine Vasopressin.
Sensitivität 7 pg/mL
Produktmerkmale EasyTest Competitive ELISA is a simple and rapid technology for the quantitation of antigen in a range of sample matrices. The whole process takes 2.5 hours with high accuracy and precision.
Bestandteile
  • Assay plate (12x8 coated Microwells)-1
  • Primary antibody-2x 6μL
  • Standard peptide -2x 10μL
  • Biotinylated peptide-2x 10μL
  • HRP-Streptavidin concentrate-1x 80μL
  • Assay diluent (5x concentrate)-1 x15mL
  • Wash buffer (20x concentrate)-1x 25 mL
  • TMB substrate-1x12 mL
  • Stop solution-1x8 mL
  • Adhensive strip-2
  • Instruction manual-1.
Benötigtes Material 1. Distilled or deionized water, 2.Precision pipettes, with disposable plastic tips, 3.Beakers, flasks, cylinders necessary for preparation of reagents, 4.Microplate washing device (multichannel pipette or automated microplate washer), 5.Microplate shaker, 6.Microplate reader capable of reading at 450 nm.
Andere Bezeichnung Vasopressin (AVP ELISA Kit Abstract)
Hintergrund Vasopressin, also known as arginine vasopressin (AVP), antidiuretic hormone (ADH), is aneurohypophysial hormone found in most mammals. It is derived from a preprohormone precursor that is synthesized in the hypothalamus and stored in vesicles at the posterior pituitary. Most of it is stored in the posterior pituitary to be released into the bloodstream. However, some AVP may also be released directly into the brain, and plays an important role in social behavior, sexual motivation and pair bonding, and maternal responses to stress.
Molekulargewicht 1084 Da
Gen-ID 280728
NCBI Accession NP_789824
UniProt P01180
Pathways cAMP Metabolic Process
Applikationshinweise Optimal working dilution should be determined by the investigator.
Probenmenge 50 μL
Testdauer 2 h
Plattentyp Pre-coated
Protokoll The immunoplate in this kit is pre-coated with secondary antibody and the nonspecific binding sites are blocked. The secondary antibody can bind to the Fc fragment of the vasopressin antibody whose Fab fragment will be competitively bound by both biotinylated vasopressin peptide and peptide standard or targeted peptide in samples. The biotinylated vasopressin peptide interacts with streptavidin-horseradish peroxidase (SA-HRP) which catalyzes TMB substrate solution. The intensity of the yellow is directly proportional to the amount of biotinylated vasopressin peptide SA-HRP complex but inversely proportional to the amount of the peptide in standard solutions or samples. A standard curve of known Vasopressin concentration can be established accordingly. The unknown Vasopressin concentration in samples can be determined by extrapolation to this standard curve.
Aufbereitung der Reagenzien
  1. Assay diluent: Dilute the concentrated assay diluent 1:5 with distilled water (e.g. 10 mL plus 40 mL). 2. Wash buffer: Dilute the concentrated wash buffer 1:20 with distilled water (e.g. 20 mL plus 380 mL). 3.Anti-Vasopressin Antibody: Briefly centrifuge the vial before use. Add 1494 μL of 1x assay diluent to the vial, mix thoroughly. 4. Standard peptide: Briefly spin standard vial before use. Add 490 μL 1x Assay Diluent to prepare a 10 ng/mL standard, gently vortex to mix, this is standard #1. Label 5 tubes #2 through #6. Pipette 150 μL 1x assay diluent into tube #2, 300 μL 1x assay buffer into tube #3, 700 μL 1x assay diluent into tubes #4 through #6. Take 150 μL from standard #1 and add to tube #2, Mix thoroughly. Add 100 μL from tube #2 to tube #3, Mix thoroughly. Continue this for tubes #4 through #6. 5.Biotinylated peptide: Briefly centrifuge the vial before use. Add 1490 μL of 1x assay diluent to make the final concentration enough for 60 wells. 6. Streptavidin-HRP: The HRP-Streptavidin concentrate should be diluted 160- fold with 1x assay diluent.
Probennahme Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4 °C before centrifugation for 15 minutes at 1000 x g. Remove serum and assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
Plasma: Collect plasma using EDTA, or citrate or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 x g at 2-8 °C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20 °C or -80 °C. Avoid repeated freeze-thaw cycles.
Aufbereitung der Proben

Levels of the target protein may vary among different specimens. Optimal dilution factors for each sample must be determined by the investigator,The dilution scheme is only suggestion: the recommended dilution for serum and plasma is 1:2.

Testdurchführung
  1. All reagents must be brought to room temperature (18-25 °C) prior to use 2. Prepare all reagents, primary anytibody, standard peptide, biotinylated peptide and samples as directed in the respective sections. 3.Determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4 °C. 4. Pipette 25 μL of anti-Vasopressin antibody into all wells, 75 μL of 1x assay diluent into the blank wells, 50 μL of 1x assay diluent into the Bo (0 ng/mL standard) wells, 50 μL of Standards or samples to the appropriate wells ,then pipette 25 μL of biotinylated peptide into each well except the Blank wells, cover the plate with adhensive strip. Incubate for 1 hour at room temperature with gentle shaking. 5.Decant or aspirate contents of wells. Wash wells by filling with at least 300 μL/well prepared wash buffer followed by decanting/aspirating. Soak wells in wash buffer for 30 seconds to 1 minute for each wash. Repeat wash 3 times for a total of 4 washes. After the last wash, blot plate on absorbent paper to remove residual buffer. Thorough washing at this step is very important, complete removal of liquid is required for proper performance. 6.Pipette 100 μL of diluted streptavidin-HRP solution to each well, Seal the plate, ncubate for 45 min at room temperature with gentle shaking. 7. wash plate as step 5. 8. Pipette 100 μL of TMB Substrate Solution to each well. Incubate plate for 15 minutes at room temperature in the dark with gentle shaking. 9. Add 50μL of Stop Solution to each well, gently tap the plate to ensure thorough mixing. 10. Determine the optical density of each well within 15 minutes using a microplate reader set to 450nm. If wavelength correction is available, set to 570nm. Subtract readings at 570nm from the readings at 450nm. This subtraction will correct for optical imperfections in the plate.
Ergebnisberechnung
  1. Average the duplicate readings for each standard and sample and subtract the average blank optical density. 2.Plot the standard curve using SigmaPlot software (or other software which can perform four-parameter logistic regression models), with standard concentration on the x-axis and percentage of absorbance (see calculation below) on the y-axis. Draw the best-fit curve through the standard points. Percentage absorbance = (B - blank OD)/ (B o - blank OD) *100 Where B = OD of sample or standard and Bo = OD of zero standard (total binding). 3. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
Testpräzision intra CV<10%, inter CV <15%
Beschränkungen Nur für Forschungszwecke einsetzbar
Buffer 0.05 % Proclin 300
Konservierungsmittel ProClin
Vorsichtsmaßnahmen The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.
Lagerung 4 °C/-20 °C
Bilder des Herstellers
ELISA image for Arginine Vasopressin (AVP) ELISA Kit (ABIN2506989) Arginine Vasopressin (AVP) ELISA Kit
Haben Sie etwas anderes gesucht?