EGFR Kit ELISA

N° du produit ABIN624965

Détail du produit EGFR Kit ELISA

Antigène: EGFR (Epidermal Growth Factor Receptor (EGFR))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 4-2000 pg/mL

Seuil minimal de détection: 4 pg/mL

Application: ELISA

Fonction: Human EGFR 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 any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensibilité: < 4 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 samples100 - 500 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: If your samples need to be diluted, Assay Diluent A should be used for dilution of serum/plasma samples. 1x Assay Diluent B should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 100-500 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.
   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 (Assay Diluent B should be diluted 5-fold with deionized or distilled water, for cell culture medium and urine) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL EGFR standard from the vial of Item C, into a tube with 960 µL Assay Diluent A or 1x Assay Diluent B to prepare a 2,000 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 . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 200myl 200 µL 200 µL 200 µL 200 µL 40 µL standard + 960 µL 2000 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
   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 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 final 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 A Human EGFR 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 Human EGFR concentration (pg/mL) O D =4 50 n m 0.001 0.01 0.1 1 10 1 10 100 1,000 10,000
Sensitivity: The minimum detectable dose of EGFR is typically less than 4 pg/mL.
Recovery: Recovery was determined by spiking various levels of human EGFR into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 105.4 94-116 Plasma 107.2 96-119 Cell culture media 85.65 72-96
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 98 97 92 Range ( %) 85-106 83-105 81-102 1:4 Average % of Expected 94 93 96 Range ( %) 83-104 82-103 85-105
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 EGFR Kit ELISA (ABIN624965)

Montermini, Meehan, Garnier, Lee, Lee, Guha, Al-Nedawi, Rak: "Inhibition of oncogenic epidermal growth factor receptor kinase triggers release of exosome-like extracellular vesicles and impacts their phosphoprotein and DNA content." dans: The Journal of biological chemistry, Vol. 290, Issue 40, pp. 24534-46, (2015) (PubMed).

Maramotti, Paci, Miccichè, Ciarrocchi, Cavazza, De Bortoli, Vaghi, Formisano, Canovi, Sgarbi, Bongarzone: "Soluble epidermal growth factor receptor isoforms in non-small cell lung cancer tissue and in blood." dans: Lung cancer (Amsterdam, Netherlands), Vol. 76, Issue 3, pp. 332-8, (2012) (PubMed).

Bhuvaneswari, Yuen, Chee, Olivo: "Antiangiogenesis agents avastin and erbitux enhance the efficacy of photodynamic therapy in a murine bladder tumor model." dans: Lasers in surgery and medicine, Vol. 43, Issue 7, pp. 651-62, (2012) (PubMed).

Détail du antigène

Antigène: EGFR (Epidermal Growth Factor Receptor (EGFR))

Autre désignation: EGFR (EGFR Produits)

Synonymes: C-erb Kit ELISA, CG10079 Kit ELISA, D-EGFR Kit ELISA, D-Egf Kit ELISA, DEGFR Kit ELISA, DER Kit ELISA, DER flb Kit ELISA, DER/EGFR Kit ELISA, DER/faint little ball Kit ELISA, DER/top Kit ELISA, DER/torpedo Kit ELISA, DER1 Kit ELISA, DEgfr Kit ELISA, Degfr Kit ELISA, Der Kit ELISA, DmHD-33 Kit ELISA, Dmel\\CG10079 Kit ELISA, EFG-R Kit ELISA, EGF-R Kit ELISA, EGFR Kit ELISA, EGFr Kit ELISA, EGfr Kit ELISA, EK2-6 Kit ELISA, Egf Kit ELISA, Egf-r Kit ELISA, EgfR Kit ELISA, El Kit ELISA, Elp Kit ELISA, Elp-1 Kit ELISA, Elp-B1 Kit ELISA, Elp-B1RB1 Kit ELISA, HD-33 Kit ELISA, TOP Kit ELISA, Torpedo/DER Kit ELISA, Torpedo/Egfr Kit ELISA, c-erbB Kit ELISA, d-egf-r Kit ELISA, dEGFR Kit ELISA, dEGFR1 Kit ELISA, dEgfr Kit ELISA, der Kit ELISA, egfr Kit ELISA, flb Kit ELISA, l(2)05351 Kit ELISA, l(2)09261 Kit ELISA, l(2)57DEFa Kit ELISA, l(2)57EFa Kit ELISA, l(2)57Ea Kit ELISA, mor1 Kit ELISA, top Kit ELISA, top/DER Kit ELISA, top/flb Kit ELISA, torpedo/Egfr Kit ELISA, torpedo/egfr Kit ELISA, EGFR12 Kit ELISA, EGFR15 Kit ELISA, egfr1 Kit ELISA, Erbb2 Kit ELISA, ERBB Kit ELISA, ERBB1 Kit ELISA, HER1 Kit ELISA, PIG61 Kit ELISA, mENA Kit ELISA, ErbB-1 Kit ELISA, Errp Kit ELISA, 9030024J15Rik Kit ELISA, AI552599 Kit ELISA, Erbb Kit ELISA, Errb1 Kit ELISA, Wa5 Kit ELISA, wa-2 Kit ELISA, wa2 Kit ELISA, epidermal growth factor receptor Kit ELISA, Epidermal growth factor receptor Kit ELISA, epidermal growth factor receptor a (erythroblastic leukemia viral (v-erb-b) oncogene homolog, avian) Kit ELISA, EGFR Kit ELISA, Egfr Kit ELISA, egfra Kit ELISA, egfr1 Kit ELISA, LOC5564544 Kit ELISA

Sujet: The Human EGFR ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human EGFR in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human EGFR coated on a 96-well plate. Standards and samples are pipetted into the wells and EGFR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human EGFR 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 EGFR 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: 1956

UniProt: P00533

Pathways: Signalisation NF-kappaB, Signalisation RTK, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Stem Cell Maintenance, Hepatitis C, Positive Regulation of Response to DNA Damage Stimulus, Interaction of EGFR with phospholipase C-gamma, Thromboxane A2 Receptor Signaling, EGFR Downregulation, S100 Proteins