Tumor Necrosis Factor Receptor Superfamily, Member 11b (TNFRSF11B) ELISA Kit

Details for Product No. ABIN2748451, Supplier: Log in to see
Antigen
  • OCIF
  • OPG
  • TR1
  • Opg
  • ocif
  • opg
  • tr1
  • TNF receptor superfamily member 11b
  • tumor necrosis factor receptor superfamily, member 11b (osteoprotegerin)
  • TNF receptor superfamily member 11B
  • tumor necrosis factor receptor superfamily member 11b S homeolog
  • tumor necrosis factor receptor superfamily, member 11b
  • TNFRSF11B
  • Tnfrsf11b
  • tnfrsf11b.S
Reactivity
Mouse (Murine)
Alternatives
Kits with alternative reactivity to:
26
21
13
9
8
4
3
3
2
2
1
1
1
Method Type
Sandwich ELISA
Detection Range
1-1200 pg/mL
Minimum Detection Limit
1 pg/mL
Application
ELISA
Options
Supplier
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Supplier Product No.
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Purpose Mouse Osteoprotegerin (TNFRSF11B) ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Plasma, Cell Culture Supernatant, Serum
Analytical Method Quantitative
Detection Method Colorimetric
Specificity This ELISA kit shows no cross-reactivity with the following cytokines tested: Mouse CD30, L CD30, T CD40, CRG-2, CTACK, CXCL16, Eotaxin , Eotaxin-2, Fas Ligand, Fractalkine, GCSF, GM-CFS, IFN- gamma, IGFBP-3, IGFBP-5, IGFBP-6, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-3 Rb, IL-4, IL-5, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17, KC, Leptin R, LEPTIN(OB), LIX, L-Selectin, Lymphotactin, MCP-1, MCP- 5, M-CSF, MIG, MIP-1 alpha, MIP-1 gamma, MIP-2, MIP-3 beta, MIP-3 alpha, PF-4, PSelectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF-alpha, TNF RI, TNF RII, TPO, VCAM-1, VEGF.
Sensitivity 1 pg/mL
Characteristics
  • 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
Components
  • 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
Material not included
  • 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
Alternative Name Osteoprotegerin (TNFRSF11B ELISA Kit Abstract)
Background Tumor necrosis factor receptor superfamily member 11B (Osteoclastogenesis inhibitory factor) (Osteoprotegerin)
Gene ID 18383
UniProt O08712
Research Area Atherosclerosis, Extracellular Matrix, Cytokines, Innate Immunity, Cardiovascular
Application Notes Recommended Dilution for serum and plasma samples20 - 200 fold
Sample Volume 100 μL
Plate Pre-coated
Protocol
  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.
Reagent Preparation
  1. Bring all reagents and samples to room temperature (18 - 25°C) before use. 2. Sample dilution: Assay Diluent A (Item D) is used for dilution of serum/plasma samples, Assay Diluent B (Item E) is used for dilution of cell culture supernatants. 3. Assay Diluent B should be diluted 5-fold with deionized or distilled water before use. 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 supernates) into Item C vial to prepare a 50 ng/ml standard. Dissolve the powder thoroughly by a gentle mix. Add 20 µl OPG standard (50 ng/ml) from the vial of Item C, into a tube with 813.3 µl Assay Diluent A or 1x Assay Diluent B to prepare a 1200 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. Gently vortex to mix. 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 10,000-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 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 100 µl of prepared 100-fold diluted solution into a tube with 10 ml 1x Assay Diluent B to prepare a final 10,000 fold diluted HRP-Streptavidin solution.
Assay Procedure
  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.
Calculation of Results

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.

Restrictions For Research Use only
Storage -20 °C
Storage Comment 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.
Expiry Date 6 months
Supplier Images
ELISA image for Tumor Necrosis Factor Receptor Superfamily, Member 11b (TNFRSF11B) ELISA Kit (ABIN2748451) Tumor Necrosis Factor Receptor Superfamily, Member 11b (TNFRSF11B) ELISA Kit
Product cited in: Carbonneau, Despars, Beaudry, Benabdallah, Bouhanik, Dépôt, Moreau, Beauséjour: "Heterotopic bone formation derived from multipotent stromal cells is not inhibited in aged mice." in: Cytotherapy, Vol. 16, Issue 8, pp. 1073-9, 2014 (PubMed).

Hartwich, Orr, Ng, Spence, McLaughlin, Furman, McGregor, Davidoff: "Rapamycin increases neuroblastoma xenograft and host stromal derived osteoprotegerin inhibiting osteolytic bone disease in a bone metastasis model." in: Journal of pediatric surgery, Vol. 48, Issue 1, pp. 47-55, 2013 (PubMed).

Celik, Gur, Akdağ, Akpolat, Guven, Celik, Sarac, Otçu: "The effects of long-term exposure to extremely low-frequency magnetic fields on bone formation in ovariectomized rats." in: Bioelectromagnetics, Vol. 33, Issue 7, pp. 543-9, 2012 (PubMed).

Chanda, Isayeva, Kumar, Siegal, Szafran, Zinn, Reddy, Ponnazhagan: "Systemic osteoprotegerin gene therapy restores tumor-induced bone loss in a therapeutic model of breast cancer bone metastasis." in: Molecular therapy : the journal of the American Society of Gene Therapy, Vol. 16, Issue 5, pp. 871-8, 2008

Chanda, Isayeva, Kumar, Siegal, Szafran, Zinn, Reddy, Ponnazhagan: "Systemic osteoprotegerin gene therapy restores tumor-induced bone loss in a therapeutic model of breast cancer bone metastasis." in: Molecular therapy : the journal of the American Society of Gene Therapy, Vol. 16, Issue 5, pp. 871-8, 2008 (PubMed).

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