IL1RN Kit ELISA

N° du produit ABIN1980008

Détail du produit IL1RN Kit ELISA

Antigène: IL1RN (Interleukin 1 Receptor Antagonist (IL1RN))

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 2-2000 pg/mL

Seuil minimal de détection: 2 pg/mL

Application: ELISA

Fonction: Mouse IL-1 Ra (IL-1 F3) ELISA Kit for cell culture supernatants, Heparin and/or EDTA treated plasma, and serum samples. Citrate is not recommended.

Type d'échantillon: Serum, Plasma, Cell Culture Supernatant

Analytical Method: Quantitative

Specificité: 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.

Sensibilité: < 2 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

Indications d'application: Recommended Dilution for serum and plasma samples5 - 50 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: 1x Assay Diluent D (Item K) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of cell culture supernates. Suggested dilution for normal serum/plasma: 5-50 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 and Assay Diluent D 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 1x Assay Diluent D (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 IL-1ra standard (50 ng/mL) from the vial of Item C, into a tube with 480 µL 1x Assay Diluent D or 1x Assay Diluent B to prepare a 2,000 pg/mL standard solution. Pipette 400 µL 1x Assay Diluent D or 1x Assay Diluent B into each tube. Use the 2,000 pg/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Assay Diluent D or 1x Assay Diluent B serves as the zero standard (0 ng/mL). The 2,000 pg/mL standard in 1x Assay Diluent B may be saturated, we recommend to start from 666.7 pg/mL for 1x Assay Diluent B Standard curve. 20 µL standard + 480 µL 200 µL 200 µL 200 µL 200 µL 200 µL 200myl
   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. 2,000 666.7 222.2 74.07 24.69 8.23 2.74 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
   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 300-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 40 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a 300-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 D Mouse IL-1ra concentration (pg/mL) O D =4 50 n m 0.001 0.01 0.1 1 10 1 10 100 1,000 10,000 Assay Diluent B Mouse IL-1ra concentration (pg/mL) O D =4 50 n m 0.1 1 10 0 1 10 100 1,000
Sensitivity: The minimum detectable dose of IL-1ra is typically less than 2 pg/mL.
Recovery: Recovery was determined by spiking various levels of mouse IL-1ra into mouse serum, plasma (EDTA and Heparin) and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 75.89 67-84 Plasma 93.96 71-111 Cell culture media 102.0 94-103
Linearity: Sample Type Serum Plasma Cell Culture Media (EDTA and Heparin) 1:2 Average % of Expected 75.78 76.49 99.32 Range ( %) 68-85 68-84 91-106 1:4 Average % of Expected 74.56 75.28 86.83 Range ( %) 67-84 67-83 69-104
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 IL1RN Kit ELISA (ABIN1980008)

Singla, Singla, Abdelli, Glass: "Fibroblast growth factor-9 enhances M2 macrophage differentiation and attenuates adverse cardiac remodeling in the infarcted diabetic heart." dans: PLoS ONE, Vol. 10, Issue 3, pp. e0120739, (2015) (PubMed).

Détail du antigène

Antigène: IL1RN (Interleukin 1 Receptor Antagonist (IL1RN))

Autre désignation: IL-1ra (IL1RN Produits)

Synonymes: DIRA Kit ELISA, ICIL-1RA Kit ELISA, IL-1RN Kit ELISA, IL-1ra Kit ELISA, IL-1ra3 Kit ELISA, IL1F3 Kit ELISA, IL1RA Kit ELISA, IRAP Kit ELISA, MVCD4 Kit ELISA, IRAP1 Kit ELISA, IL1RN Kit ELISA, IL-1RA Kit ELISA, F630041P17Rik Kit ELISA, interleukin 1 receptor antagonist Kit ELISA, interleukin 1 receptor antagonist-like Kit ELISA, IL1RN Kit ELISA, Il1rn Kit ELISA, IL1RNL Kit ELISA

Sujet: The Mouse IL-1ra ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of IL-1ra in serum, plasma (collect plasma using EDTA and heparin as an anticoagulant. Citrate is not recommended for this assay). This assay employs an antibody specific for mouse IL-1ra coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-1ra present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse IL-1ra 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 IL-1ra 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: 16181

UniProt: P25085

Pathways: Signalisation NF-kappaB, Hormone Transport, Cancer Immune Checkpoints