TNFRSF18 ELISA Kit (Tumor Necrosis Factor Receptor Superfamily, Member 18)

Details for Product TNFRSF18 ELISA Kit No. ABIN624986, Anbieter: Anmelden zum Anzeigen
Antigen
  • GITR
  • TNFRSF18
  • AITR
  • Gitr
  • CD357
  • GITR-D
  • TNF receptor superfamily member 18
  • tumor necrosis factor receptor superfamily, member 18
  • TNFRSF18
  • Tnfrsf18
Reaktivität
Human
Alternativen
Kits mit alternativen Reaktivitäten:
13
7
Methodentyp
Sandwich ELISA
Detektionsbereich
15-10000 pg/mL
Untere Nachweisgrenze
15 pg/mL
Applikation
ELISA
Optionen
Hersteller
Anmelden zum Anzeigen
Hersteller Produkt- Nr.
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Verwendungszweck Human GITR (TNFRSF18) ELISA Kit for cell culture supernatants, plasma, and serum samples.
Proben Plasma, Cell Culture Supernatant, Serum
Analytische Methode Quantitative
Nachweismethode Colorimetric
Spezifität 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.
Sensitivität 15 pg/mL
Produktmerkmale
  • 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
Bestandteile
  • 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
Benötigtes Material
  • 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 Plasmide, Primers & weitere TNFRSF18 products on genomics-online (e.g. as negative or positive controls)
Antigen
Andere Bezeichnung GITR (TNFRSF18 ELISA Kit Abstract)
Hintergrund GITR (glucocorticoid induced tumor necrosis factor receptor family related gene) is a type-1 transmembrane protein of 228 amino acids belonging to the TNF and NGF receptor family of proteins. GITR is expressed in normal T-lymphocytes from thymus, spleen. Constitutive expression of a transfected GITR gene induces resistance to apoptosis induced by anti CD3 monoclonal antibodies. The human homolog of GITR is expressed in lymph node and peripheral blood leukocytes. Its expression is up-regulated in human peripheral mononuclear cells mainly after stimulation with antibodies against CD3 and CD28. The Human GITR ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human GITR in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human GITR coated on a 96-well plate. Standards and samples are pipetted into the wells and GITR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human GITR 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 GITR 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%.
Gen-ID 8784
UniProt Q9Y5U5
Applikations-hinweise Recommended Dilution for serum and plasma samples2 fold
Probenmenge 100 μL
Plattentyp Pre-coated
Protokoll
  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.
Aufbereitung der Reagenzien
  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) is used for dilution of serum/plasma samples, and Assay Diluent B (Item E) is used for dilution of culture supernatants and urine. 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, Assay Diluent B should be diluted 5-fold with deionized or distilled water before use) into Item C vial to prepare a 100 ng/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 60 µl GITR standard from the vial of Item C, into a tube with 540 µl Assay Diluent A or 1x Assay Diluent B to prepare a 10,000 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 (shown below). Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 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) and pipette up and down to mix gently before use. HRP- Streptavidin concentrate should be diluted 20,000-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 2 µl of HRP-Streptavidin concentrate into a tube with 198.0 µl 1x Assay Diluent B to prepare a 100-fold diluted HRP- Streptavidin solution (do not store the diluted solution for next day use). Mix through and then pipette 70 µl of prepared 100-fold diluted solution into a tube with 14 ml 1x Assay Diluent B to prepare a final 20,000 fold diluted HRP-Streptavidin solution.
Testdurchführung
  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 µl) 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 3. 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 3. 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.
Ergebnisberechnung

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.

Beschränkungen Nur für Forschungszwecke einsetzbar
Lagerung -20 °C
Informationen zur Lagerung 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.
Haltbarkeit 6 months
Bilder des Herstellers
ELISA image for Tumor Necrosis Factor Receptor Superfamily, Member 18 (TNFRSF18) ELISA Kit (ABIN624986) Tumor Necrosis Factor Receptor Superfamily, Member 18 (TNFRSF18) ELISA Kit
Produkt verwendet in: Gan, Feng, Gu, Tan, Sun, Lv, Zhang: "Correlation of increased blood levels of GITR and GITRL with disease severity in patients with primary Sjögren's syndrome." in: Clinical & developmental immunology, Vol. 2013, pp. 340751, 2013 (PubMed).

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