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Human METTL4 Kit Elisa (A326727)

Standard Curve - Human METTL4 ELISA Kit (A326727) - Antibodies.com
$616
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Produktname
Human METTL4 ELISA Kit
Beschreibung
Human METTL4 ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human METTL4 in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples.
Assay-Typ
Sandwich (quantitative)
Assay-Prinzip
Human METTL4 ELISA Kit (A326727) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human METTL4 in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. An antibody specific for METTL4 has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the METTL4 present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with Biotinylated Anti-METTL4 Antibody, which binds the captured METTL4 present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Streptavidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of METTL4 captured in each well. The concentration of METTL4 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
Plattform
Pre-coated Microplate (12 x 8 well strips)
Nachweismethode
Colorimetric
Instrument
Colorimetric Microplate Reader
Probentyp
Serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples.
Sensitivität
0.094 ng/ml
Detektionsbereich
0.156-10 ng/ml
Testdauer
4 hours
Reaktivität
Human
Recovery
Sample Type n Range Average
Serum 10 85% - 102% 94%
EDTA Plasma 10 86% - 104% 93%
Heparin Plasma 10 92% - 99% 95%
Linearity
Sample Type n 1:2 1:4 1:8
Serum 10 88-103% 83-101% 83-98%
EDTA Plasma 10 85-105% 92-99% 83-97%
Heparin Plasma 10 88-98% 83-99% 81-100%
Precision
Intra-assay CV: < 8%, Inter-assay CV: < 10%
Allgemeine Hinweise
Information online is representative. Always refer to the Product Manual inside the kit.
Lagerung
Store at +4°C. Do not use past expiration date!
Synonyms
HsT661, Methyltransferase like 4, Methyltransferase-like protein 4, METL4_HUMAN, METTL 4
Haftungsausschluss
Dieses Produkt ist nur für Forschungszwecke bestimmt. Es ist nicht für diagnostische oder therapeutische Zwecke bestimmt.
Kit Components
Item Quantity Storage
Pre-Coated 96 Well Microplate 12 x 8 Well Strips +4°C
Lyopholized Standard 2 Vials +4°C
Sample Dilution Buffer 20ml +4°C
Biotinylated Detection Antibody 120µl +4°C
Antibody Dilution Buffer 10ml +4°C
HRP-Streptavidin Conjugate 120µl +4°C
SABC Dilution Buffer 10ml +4°C
TMB Substrate 10ml +4°C
Stop Solution 10ml +4°C
Wash Buffer (25X) 30ml +4°C
Plate Sealers 5 Adhesive Strips -
Foil Pouch 1 Zip-Sealed Pouch -
Required Materials
The following materials are not included in the kit, but are required to run this assay:
  • Microplate reader capable of measuring absorbance at 450nm ± 10nm.
  • Squirt bottle, multichannel pipette reservoir, or automated microplate washer.
  • Graph paper or computer software capable of generating logarithmic functions.
  • Test tubes and Eppendorf tubes to prepare standards and samples.
  • Multi- and single-channel pipettes and sterile pipette tips.
  • Deionized or distilled water.
  • Absorbent paper towels.
  • 37°C incubator.
Important Notes
  • Sample concentrations should be estimated before being used in the assay and a proper dilution factor should be selected to make the diluted target protein concentration fall in the optimal detection range of this kit.
  • Sample matrix components will interfere with the test results. All samples must be diluted at least 1:2 with Sample Dilution Buffer.
  • Hemolysis can greatly impact the validity of test results. Take care to minimize hemolysis.
Bring all reagents and samples to room temperature before use.
We recommend that all standards and samples be assayed in duplicate.
Step 1
Prepare all reagents, samples, and working standards as directed in the previous sections.
Step 2
Determine the number of microplate strips required for the assay and insert them into the plate frame. Unused strips should be stored in the foil pouch at +4°C.
Step 3
Add 100μl of standard solutions to the standard wells. Note: A plate layout is provided to record standards and samples assayed.
Step 4
Add 100μl of samples to the sample wells. Note: All samples must be properly diluted in order to produce sample values within the dynamic range of the assay. All samples must be diluted at least 1:2 with Sample Dilution Buffer.
Step 5
Cover with an adhesive plate sealer provided and incubate at 37°C for 90 minutes.
Step 6
Remove the plate sealer, aspirate the liquid from the plate, and wash the plate two times with Wash Buffer. Soak wells with 350μl of Wash Buffer for 1-2 minutes each time. Note: Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Blot the plate onto paper towels or other absorbent material. Do not let the wells dry completely at any time!
Step 7
Add 100μl of Biotinylated Detection Antibody working solution to each well.
Step 8
Cover with a new adhesive plate sealer and incubate at 37°C for 60 minutes.
Step 9
Remove the plate sealer, aspirate the liquid from the plate, and wash the plate three times with Wash Buffer. Soak wells with 350μl of Wash Buffer for 1-2 minutes each time.
Step 10
Add 100μl of HRP-Streptavidin Conjugate (SABC) working solution to each well.
Step 11
Cover with a new adhesive plate sealer and incubate at 37°C for 60 minutes.
Step 12
Remove the plate sealer, aspirate the liquid from the plate, and wash the plate five times with Wash Buffer. Soak wells with 350μl of Wash Buffer for 1-2 minutes each time.
Step 13
Add 90μl of TMB Substrate to each well. Cover with a new adhesive plate sealer and incubate at 37°C in the dark for 10-20 minutes. Note: This incubation time is for reference only, the optimal reaction time should be determined by the end-user and can be shortened or extended according to the actual color change. The reaction may be terminated when a gradient is apparent in the standard wells. Do not exceed 30 minutes!
Step 14
Add 50μl of Stop Solution to each well. The color in the wells should change from blue to yellow immediately. Note: If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.
Step 15
Determine the optical density (O.D. value) of each well immediately using a microplate reader set to 450 nm.
Standard Curve - Human METTL4 ELISA Kit (A326727) - Antibodies.comBild Vergrößern
Figure 1: Standard Curve - Human METTL4 ELISA Kit (A326727)
Representative standard curve for Human METTL4 ELISA Kit (A326727).
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