IL6ST Kit ELISA (Interleukin 6 Signal Transducer (Gp130, Oncostatin M Receptor))

Details for Product IL6ST ELISA Kit No. ABIN625085, Fournisseur: Connectez-vous pour afficher
Antigène
  • CD130
  • CDW130
  • GP130
  • IL-6RB
  • PEPN
  • 5133400A03Rik
  • AA389424
  • BB405851
  • D13Ertd699e
  • gp130
  • Ac1055
  • Gp130
  • Il-6rb
  • IL-6
  • interleukin 6 signal transducer
  • alanyl aminopeptidase, membrane
  • IL6ST
  • ANPEP
  • Il6st
  • il6st
Reactivité
Humain
Alternatives
Kits with alternative reactivity to:
13
12
5
Type de méthode
Sandwich ELISA
Gamme de detection
4-6000 pg/mL
Seuil minimal de détection
4 pg/mL
Application
ELISA
Options
Fournisseur
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N° du produit (Fournisseur)
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Fonction Human gp130 ELISA Kit for cell culture supernatants, plasma, and serum samples.
Type d'échantillon Plasma, Cell Culture Supernatant, Serum
Analytical Method Quantitative
Méthode de détection Colorimetric
Specificité This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
Sensibilité < 4 pg/mL
Attributs du produit
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data
Ingrédients
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Matériel non inclus
  • Distilled or deionized water
  • Precision pipettes to deliver 2 μL to 1 μL volumes
  • Adjustable 1-25 μL pipettes for reagent preparation
  • 100 μL and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Plasmids, Primers & others Plasmids, Primers & others IL6ST products on genomics-online (e.g. as negative or positive controls)
Antigène Interleukin 6 Signal Transducer (Gp130, Oncostatin M Receptor) (IL6ST)
Autre désignation Gp130 (IL6ST ELISA Kit Extrait)
Sujet The Human sgp-130 ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human sgp-130 in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human sgp-130 coated on a 96-well plate. Standards and samples are pipetted into the wells and sgp-130 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human sgp-130 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of sgp-130 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
ID gène 3572
UniProt P40189
Pathways Signalistation JAK/STAT, Cellular Glucan Metabolic Process, Autophagy, Smooth Muscle Cell Migration
Indications d'application Recommended Dilution for serum and plasma samples100 - 1,000 fold
Volume d'échantillon 100 μL
Plaque Pre-coated
Protocole
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 μL of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 μL of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 μL of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 μL of Stop Solution to each well.
  11. Read at 450 nm immediately.
Préparation des réactifs
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
    2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 100-1000 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
    3. Assay Diluent B should be diluted 5-fold with deionized or distilled water.
    4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 80 µL sgp-130 standard from the vial of Item C, into a tube with 586.7 µL Assay Diluent A or 1x Assay Diluent B to prepare a 6,000 pg/mL stock standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero. 200 µL 200myl 200 µL 200 µL 200 µL 200 µL 80 µL standard + 586.7 µL 6,000 2,000 666.7 222.2 74.07 24.69 8.23 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
    5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
    6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
    7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) before use. HRP-Streptavidin concentrate should be diluted 160-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 75 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluen B to prepare a 160-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
Procédure de l'essai
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
    2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
    3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. 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. Invert the plate and blot it against clean paper towels.
    4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
    5. Discard the solution. Repeat the wash as in step
    6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
    7. Discard the solution. Repeat the wash as in step
    8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
    9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calcul des résultats

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent A Human sgp-130 concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 . Assay Diluent B Human sgp-130 concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of sgp-130 is typically less than 4 pg/mL.
Recovery: Recovery was determined by spiking various levels of human sgp-130 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 95.37 84-105 Plasma 94.66 82-104 Cell culture media 97.35 87-108
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 94 95 96 Range ( %) 85-103 86-105 87-106 1:4 Average % of Expected 95 96 96 Range ( %) 86-106 88-105 89-107 1:8 Average % of Expected 97 96 97 Range ( %) 87-107 89-107 90-106
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Précision du teste Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
Restrictions For Research Use only
Conseil sur la manipulation Avoid repeated freeze-thaw cycles.
Stock -20 °C
Stockage commentaire The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Date de péremption 6 months
Images (Fournisseur)
ELISA image for Interleukin 6 Signal Transducer (Gp130, Oncostatin M Receptor) (IL6ST) ELISA Kit (ABIN625085) Interleukin 6 Signal Transducer (Gp130, Oncostatin M Receptor) (IL6ST) ELISA Kit
Produit citée dans: Gao, Camous, Lu, Lim, Larbi, Ng: "Novel inflammatory markers associated with cognitive performance: Singapore Longitudinal Ageing Studies." dans: Neurobiology of aging, Vol. 39, pp. 140-6, 2016 (PubMed). (Échantillon (espèces): Human). Détails: Sample: Plasma (Cognitive Performance)

Lu, Zhu, Jiang, Sun, Jia, Qian, Li, Ma: "Matrine increases NKG2D ligand ULBP2 in K562 cells via inhibiting JAK/STAT3 pathway: a potential mechanism underlying the immunotherapy of matrine in leukemia." dans: American journal of translational research, Vol. 7, Issue 10, pp. 1838-49, 2015 (PubMed).

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