Pecora TNF alpha Kit Elisa (A7658)

Spese di spedizione
Tempi di consegna
7-10 Giorni lavorativi
Telefono
+1 (314) 370-6046
Lun - Ven, 8:00 - 16:00 AST
E-mail
orders@antibodies.com
Nome
Sheep TNF alpha ELISA Kit
Descrizione
Sheep TNF alpha ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of sheep TNF alpha in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Tipo di kit
Sandwich (quantitative)
Principio di kit
Sheep TNF alpha ELISA Kit (A7658) employs the sandwich enzyme immunoassay technique for the quantitative measurement of sheep TNF alpha in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. An antibody specific for TNF alpha has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the TNF alpha 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-TNF alpha Antibody, which binds the captured TNF alpha present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Avidin 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 TNF alpha captured in each well. The concentration of TNF alpha can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
Piattaforma
Pre-coated Microplate (12 x 8 well strips)
Tipo di rilevamento
Colourmetric
Instrument
Colorimetric Microplate Reader
Tipo di campione
Serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Sensibilità
7.1 pg/ml
Campo di rilevamento
15.6-1000 pg/ml
Tempo del kit
4h 30m
Reattività
Sheep
Recovery
Sample Type n Range Average
Serum 5 80% - 102% 91%
EDTA Plasma 5 81% - 100% 90%
Heparin Plasma 5 80% - 89% 84%
Linearity
Sample Type 1:2 1:4 1:8 1:16
Serum (n=5) 87-91% 87-107% 74-101% 92-97%
EDTA Plasma (n=5) 90-105% 84-101% 90-101% 79-108%
Heparin Plasma (n=5) 84-95% 92-105% 82-105% 89-91%
Precision
Intra-assay CV: < 8%, Inter-assay CV: < 10%
Note generali
Information online is representative. Always refer to the Product Manual inside the kit.
Conservazione
Store kit components at +4°C or -20°C as instructed. Do not use past expiration date!
Sinonimi
APC1, APC1 protein, Cachectin, DIF, Differentiation inducing factor, Macrophage cytotoxic factor, soluble form, Tnf, TNF a, TNF macrophage derived, TNF monocyte derived, TNF superfamily member 2 , TNF superfamily, member 2, TNF, macrophage derived, TNF, monocyte derived, TNF-a, TNF-alpha, TNFA, TNFA_HUMAN, TNFSF2, Tumor necrosis factor, Tumor necrosis factor (TNF superfamily member 2) , Tumor necrosis factor alpha, Tumor necrosis factor ligand superfamily member 2, Tumor Necrosis Factor, Membrane Form, Tumor necrosis factor, soluble form
Dichiarazione di non responsabilità
Questo prodotto è solo per uso di ricerca. Non è inteso per uso diagnostico o terapeutico.
Kit Components
Item Quantity Storage
Pre-Coated 96 Well Microplate 12 x 8 Well Strips -20°C
Lyopholized Standard 2 Vials -20°C
Detection Solution A 120μl -20°C
Detection Solution B 120µl -20°C
Wash Buffer (30X) 20ml +4°C
Sample Dilution Buffer 45ml -20°C
TMB Substrate 9ml +4°C
Stop Solution 6ml +4°C
Plate Sealers 5 Adhesive Strips -
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.
  • Hemolysis can greatly impact the validity of test results. Take care to minimize hemolysis.
  • Samples should be brought to room temperature before starting the assay.
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 a foil pouch at -20°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.
Step 5
Cover with an adhesive plate sealer provided and incubate at 37°C for 2 hours.
Step 6
Remove the plate sealer and aspirate the liquid from the plate.
Step 7
Add 100μl of working Detection Solution A 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 300μl 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 10
Add 100μl of working Detection Solution B 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 300μl of Wash Buffer for 1-2 minutes each time.
Step 13
Add 90μl of TMB Substrate to each well. Cover with a new plate sealer and incubate at 37°C in the dark for 15-25 minutes (do not exceed 30 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.
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.

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