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TNF alpha Kit ELISA

Small Sample TNF alpha Reactivité: Souris Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
N° du produit ABIN4948107
  • Antigène Voir toutes TNF alpha Kits ELISA
    TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))
    Reactivité
    • 14
    • 12
    • 10
    • 5
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Souris
    Méthode de détection
    Colorimetric
    Type de méthode
    Sandwich ELISA
    Gamme de detection
    15.6 pg/mL - 1000 pg/mL
    Seuil minimal de détection
    15.6 pg/mL
    Application
    ELISA
    Fonction
    Mini Samples ELISA Kit for Tumor Necrosis Factor Alpha (TNFa)
    Type d'échantillon
    Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
    Analytical Method
    Quantitative
    Specificité

    This assay has high sensitivity and excellent specificity for detection of Mini Samples Tumor Necrosis Factor Alpha (TNFa).
    No significant cross-reactivity or interference between Mini Samples Tumor Necrosis Factor Alpha (TNFa) and analogues was observed.

    Sensibilité
    5.9 pg/mL
    Classe de qualité
    Small Sample
    Ingrédients
    • 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 x concentrate)
    • Instruction manual
    Matériel non inclus
    • 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
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  • Indications d'application
    • 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.
    Commentaires

    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.

    Volume d'échantillon
    25 μL
    Durée du test
    3 h
    Plaque
    Pre-coated
    Protocole
    The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Mini Samples Tumor Necrosis Factor Alpha (TNFa). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mini Samples Tumor Necrosis Factor Alpha (TNFa). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Mini Samples Tumor Necrosis Factor Alpha (TNFa), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Mini Samples Tumor Necrosis Factor Alpha (TNFa) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
    Préparation des réactifs
    1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
    2. Standard - Reconstitute the Standard with 0.5 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 1,000pg/mL. Prepare 7 tubes containing 0.25 mL Standard Diluent and produce a double dilution series by transferring 250 µL each. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 1,000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
    3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stockDetection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
    4. Wash Solution - Dilute 10 mL of Wash Solution concentrate (30x) with 290 mL of deionized or distilled water to prepare 300 mL of Wash Solution (1x).
    5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

    Note:

    1. Making serial dilution in the wells directly is not permitted.
    2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
    3. 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.
    4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
    5. Prepare Substrate working Solution within 15 minutes before assay.
    6. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
    7. Contaminated water or container for reagent preparation will influence the detection result.
    Préparation de l'échantillon
    • 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.
    Procédure de l'essai
    1. Prepare all reagents, samples and standards,
    2. Add 25μL standard or sample to each well. Incubate 1 hour at 37 °C,
    3. Aspirate and add 25μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
    4. Aspirate and wash 3 times,
    5. Add 25μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
    6. Aspirate and wash 5 times,
    7. Add 25μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
    8. Add 20μL Stop Solution. Read at 450nm immediately.
    Précision du teste

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

    Restrictions
    For Research Use only
  • Précaution d'utilisation
    The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
    Conseil sur la manipulation
    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.
    Stock
    4 °C
    Stockage commentaire
    • 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.
    Date de péremption
    6 months
  • Antigène Voir toutes TNF alpha Kits ELISA
    TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))
    Autre désignation
    Tumor Necrosis Factor Alpha (TNF alpha Produits)
    Synonymes
    DIF Kit ELISA, TNF-alpha Kit ELISA, TNFA Kit ELISA, TNFSF2 Kit ELISA, RATTNF Kit ELISA, Tnfa Kit ELISA, tnf Kit ELISA, TNF-a Kit ELISA, TNFalpha Kit ELISA, Tnfsf1a Kit ELISA, TNFa Kit ELISA, cTNF Kit ELISA, Tnf-alpha Kit ELISA, tnfa-like Kit ELISA, TNF-ALPHA Kit ELISA, dif Kit ELISA, tnfa Kit ELISA, xtnf Kit ELISA, tnfsf2 Kit ELISA, tnf-alpha Kit ELISA, Cachectin Kit ELISA, tumor necrosis factor Kit ELISA, tumor necrosis factor b (TNF superfamily, member 2) Kit ELISA, tumor necrosis factor alpha Kit ELISA, tumor necrosis factor a (TNF superfamily, member 2) Kit ELISA, TNF Kit ELISA, Tnf Kit ELISA, tnf Kit ELISA, tnfb Kit ELISA, tnf-alpha Kit ELISA, LOC103694380 Kit ELISA, tnfa Kit ELISA
    UniProt
    P06804
    Pathways
    Signalisation NF-kappaB, Apoptose, Caspase Cascade in Apoptosis, Signalisation TLR, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus, Inflammasome
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