IGF2 Kit ELISA

N° du produit ABIN625133

Détail du produit IGF2 Kit ELISA

Antigène: IGF2 (Insulin-Like Growth Factor 2 (IGF2))

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Fonction: Mouse IGF-2 ELISA Kit for Cell Culture Supernatants samples.

Type d'échantillon: Cell Culture Supernatant

Analytical Method: Quantitative

Specificité: The antibody pair provided in this kit recognizes mouse IGF-II (for Cell Culture).

Sensibilité: 1500 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

Information d'application

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 (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.

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 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 %

Précision du teste: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Restrictions: For Research Use only

Stockage

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

Références pour IGF2 Kit ELISA (ABIN625133)

Dobie, Ahmed, Staines, Pass, Jasim, MacRae, Farquharson: "Increased linear bone growth by GH in the absence of SOCS2 is independent of IGF-1." dans: 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." dans: 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." dans: Southern medical journal, Vol. 58, Issue 11, pp. 1420-2 (PubMed).

Détail du antigène

Antigène: IGF2 (Insulin-Like Growth Factor 2 (IGF2))

Autre désignation: IGF-II (IGF2 Produits)

Synonymes: C11orf43 Kit ELISA, IGF-II Kit ELISA, PP9974 Kit ELISA, LOC100136550 Kit ELISA, igf2 Kit ELISA, igf2b Kit ELISA, wu:fc26e03 Kit ELISA, zgc:91781 Kit ELISA, igf-2 Kit ELISA, insigf Kit ELISA, pp9974 Kit ELISA, AL033362 Kit ELISA, Igf-2 Kit ELISA, Igf-II Kit ELISA, M6pr Kit ELISA, Mpr Kit ELISA, Peg2 Kit ELISA, IGFII Kit ELISA, RNIGF2 Kit ELISA, IGF-II-B Kit ELISA, igf2-a Kit ELISA, igf2-b Kit ELISA, insulin like growth factor 2 Kit ELISA, insulin-like growth factor II Kit ELISA, insulin-like growth factor 2a Kit ELISA, insulin like growth factor 2 S homeolog Kit ELISA, insulin-like growth factor 2 Kit ELISA, insulin-like growth factor 2 (somatomedin A) Kit ELISA, insulin like growth factor 2 L homeolog Kit ELISA, IGF2 Kit ELISA, LOC100136550 Kit ELISA, igf2a Kit ELISA, igf2.S Kit ELISA, Igf2 Kit ELISA, igf2.L Kit ELISA

Sujet: 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%.

ID gène: 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