Phone:
+1 877 302 8632
Fax:
+1 888 205 9894 (Toll-free)
E-Mail:
orders@antibodies-online.com

IGF2 ELISA Kit

IGF2 Reactivity: Mouse Colorimetric Sandwich ELISA Cell Culture Supernatant
Catalog No. ABIN625133
  • Target See all IGF2 ELISA Kits
    IGF2 (Insulin-Like Growth Factor 2 (IGF2))
    Reactivity
    • 9
    • 6
    • 4
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Mouse
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Application
    ELISA
    Purpose
    Mouse IGF-2 ELISA Kit for Cell Culture Supernatants samples.
    Sample Type
    Cell Culture Supernatant
    Analytical Method
    Quantitative
    Specificity
    The antibody pair provided in this kit recognizes mouse IGF-II (for Cell Culture).
    Sensitivity
    1500 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
    Featured
    Discover our best selling IGF2 ELISA Kit
    Top Product
    Discover our top product IGF2 ELISA Kit
  • 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: If your samples need to be diluted, Assay diluent (Item E) should be used for dilution of cell culture supernates.
      3. Assay diluent (Item E) 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 500 µL 1x Assay diluent (Item E, should be diluted 5-fold with deionized or distilled water before use) into Item C vial to prepare a 500 ng/mL stock standard. Dissolve the powder thoroughly by a gentle mix. Add 200 µL IGF-II standard from the vial of Item C, into a tube with 466.7 µL 1x Assay Diluent to prepare a 150 ng/mL standard solution. Pipette 300myl 1x Assay diluent into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay diluent serves as the zero standard (0 ng/mL). 300 µL 300 µL 300 µL 300 µL 300 µL 300myl 200 µL standard + 466.7 µL 150 75 37.5 18.8 9.4 4.7 2.3 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/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 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 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 1000-fold with 1x Assay diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 15 µL of HRP-Streptavidin concentrate into a tube with 15 ml 1x Assay Diluent to prepare a 1000-fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    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 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.
    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.
    Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent Buffer Mouse IGF-II concentration (ng/mL) O D =4 50 n m 0.1 1 10 0 10 100 1000
    Sensitivity: The minimum detectable dose of IGF-II is typically less than 1.5 ng/mL.
    Recovery: Recovery was determined by spiking various levels mouse IGF-II into cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Cell culture media 109.7 96-123
    Linearity: Sample Type Cell Culture Media 1:2 Average % of Expected 87.58 Range ( %) 77-98 1:4 Average % of Expected 85.98 Range ( %) 77-95
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Assay Precision
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Restrictions
    For Research Use only
  • Handling Advice
    Avoid repeated freeze-thaw cycles.
    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
  • Dobie, Ahmed, Staines, Pass, Jasim, MacRae, Farquharson: "Increased linear bone growth by GH in the absence of SOCS2 is independent of IGF-1." in: Journal of cellular physiology, Vol. 230, Issue 11, pp. 2796-806, (2015) (PubMed).

    Talbot, Sparks, Powell, Kahl, Caperna: "Quantitative and semiquantitative immunoassay of growth factors and cytokines in the conditioned medium of STO and CF-1 mouse feeder cells." in: In vitro cellular & developmental biology. Animal, Vol. 48, Issue 1, pp. 1-11, (2012) (PubMed).

    Wilkinson, Frankel: "The temporary hanging hip procedure: a preliminary report." in: Southern medical journal, Vol. 58, Issue 11, pp. 1420-2 (PubMed).

  • Target See all IGF2 ELISA Kits
    IGF2 (Insulin-Like Growth Factor 2 (IGF2))
    Alternative Name
    IGF-II (IGF2 Products)
    Synonyms
    C11orf43 ELISA Kit, IGF-II ELISA Kit, PP9974 ELISA Kit, LOC100136550 ELISA Kit, igf2 ELISA Kit, igf2b ELISA Kit, wu:fc26e03 ELISA Kit, zgc:91781 ELISA Kit, igf-2 ELISA Kit, insigf ELISA Kit, pp9974 ELISA Kit, AL033362 ELISA Kit, Igf-2 ELISA Kit, Igf-II ELISA Kit, M6pr ELISA Kit, Mpr ELISA Kit, Peg2 ELISA Kit, IGFII ELISA Kit, RNIGF2 ELISA Kit, IGF-II-B ELISA Kit, igf2-a ELISA Kit, igf2-b ELISA Kit, insulin like growth factor 2 ELISA Kit, insulin-like growth factor II ELISA Kit, insulin-like growth factor 2a ELISA Kit, insulin like growth factor 2 S homeolog ELISA Kit, insulin-like growth factor 2 ELISA Kit, insulin-like growth factor 2 (somatomedin A) ELISA Kit, insulin like growth factor 2 L homeolog ELISA Kit, IGF2 ELISA Kit, LOC100136550 ELISA Kit, igf2a ELISA Kit, igf2.S ELISA Kit, Igf2 ELISA Kit, igf2.L ELISA Kit
    Background
    The Mouse IGF-II ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse IGF-II in cell culture supernates. This assay employs an antibody specific for mouse IGF-II coated on a 96-well plate. Standards and samples are pipetted into the wells and IGF-II present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse IGF-II 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 IGF-II 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%.
    Gene ID
    16002
    UniProt
    P09535
    Pathways
    Hormone Activity, Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process, Regulation of Carbohydrate Metabolic Process, Activated T Cell Proliferation
You are here:
Support