CCL20 Kit ELISA

N° du produit ABIN625165

Détail du produit CCL20 Kit ELISA

Antigène: CCL20 (Chemokine (C-C Motif) Ligand 20 (CCL20))

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 3-1500 pg/mL

Seuil minimal de détection: 3 pg/mL

Application: ELISA

Fonction: Mouse MIP-3 alpha (CCL20) ELISA Kit for cell culture supernatants, plasma, and serum samples.

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

Analytical Method: Quantitative

Specificité: This ELISA kit shows no cross-reactivity with the following cytokines tested: Mouse CD30, CD30L, 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, PF-4, P-Selectin, RANTES, SCF, SDF-1 alpha, TARC, TCA-3, TECK, TIMP-1, TNF-alpha, TNF RI, TNF RII, TPO, VCAM-1, VEGF.

Sensibilité: < 3 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 samples2 - 10 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: Assay Diluent A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of cell culture supernatants. Suggested dilution for normal serum/plasma: 2-10 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 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 15 µL MIP-3 alpha standard (50 ng/mL) from the vial of Item C, into a tube with 485 µL Assay Diluent A or 1x Assay Diluent B to prepare a 1,500 pg/mL standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the 1,500 pg/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 15 µL standard + 485 µL 200 µL 200 µL 200 µL 200 µL 200 µL 200myl 1,500 500 166.7 55.56 18.52 6.17 2.06 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL 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 400-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 30 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a 400-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 A Mouse MIP-3 alpha concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B Mouse MIP-3 alpha concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.001 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of MIP-3 alpha is typically less than 3 pg/mL.
Recovery: Recovery was determined by spiking various levels of mouse MIP-3 alpha into mouse serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 83.74 74-93 Plasma 108.2 84-143 Cell culture media 80.52 72-91
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 95.87 112.0 126.4 Range ( %) 86-104 104-121 115-124 1:4 Average % of Expected 74.07 83.60 85.04 Range ( %) 68-89 70-106 75-94
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 CCL20 Kit ELISA (ABIN625165)

Shanmugam, Wang, Markand, Perry, Tawfik, Zorrilla, Ganapathy, Smith: "Sigma receptor 1 activation attenuates release of inflammatory cytokines MIP1?, MIP2, MIP3?, and IL12 (p40/p70) by retinal Müller glial cells." dans: Journal of neurochemistry, (2015) (PubMed).

Détail du antigène

Antigène: CCL20 (Chemokine (C-C Motif) Ligand 20 (CCL20))

Autre désignation: MIP-3 alpha / CCL20 (CCL20 Produits)

Synonymes: MIP-3ALPHA Kit ELISA, CCL20 Kit ELISA, si:dkey-150o13.1 Kit ELISA, CKb4 Kit ELISA, LARC Kit ELISA, MIP-3-alpha Kit ELISA, MIP-3a Kit ELISA, MIP3A Kit ELISA, SCYA20 Kit ELISA, ST38 Kit ELISA, MIP-3A Kit ELISA, MIP-3[a] Kit ELISA, Scya20 Kit ELISA, exodus-1 Kit ELISA, C-C motif chemokine ligand 20 Kit ELISA, chemokine (C-C motif) ligand 20b Kit ELISA, chemokine (C-C motif) ligand 20 Kit ELISA, CCL20 Kit ELISA, ccl20b Kit ELISA, Ccl20 Kit ELISA

Sujet: The Mouse MIP-3 alpha (Marcrophage Inflammatory Protein 3 alpha) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse MIP-3 alpha in serum, plasma and cell culture supernatants. This assay employs an antibody specific for mouse MIP-3 alpha coated on a 96-well plate. Standards and samples are pipetted into the wells and MIP-3 alpha present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse MIP-3 alpha 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 MIP-3 alpha 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: 20297

UniProt: O89093

Pathways: Phosphorylation & l'infection par le SRAS-CoV-2