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Human Amphiregulin ELISA Kit

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Antigen
Amphiregulin (AREG) ELISA Kits
  • AR
  • Sdgf
  • CRDGF
  • SDGF
  • AREG
  • AREG/SDGF
  • AREGB
  • amphiregulin
  • amphiregulin B
  • amphiregulin preproprotein-like
  • amphiregulin-like
  • Areg
  • AREG
  • AREGB
  • LOC100354550
  • LOC538751
  • LOC100050528
  • LOC100732447
Reactivity
Human
Alternatives
Kits with alternative reactivity to:
18
17
12
7
7
7
7
3
3
2
1
1
1
1
Method Type
Sandwich ELISA
Detection Range
10-4000 pg/mL
Minimum Detection Limit
10 pg/mL
Application
ELISA
Options
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Catalog No. ABIN624943
$ 476.30
Plus shipping costs $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Supplier References Details
11.926027 ABIN414351 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.6-1000 pg/mL 15.6 pg/mL Log in to see 6
11.926027 ABIN1889290 Sandwich ELISA Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA), Saliva, Milk 15.6-1000 pg/mL 15.6 pg/mL Log in to see 3
11.926027 ABIN365786 Sandwich ELISA Serum, Plasma 4.7-300 pg/mL 4.7 pg/mL Log in to see
11.926027 ABIN1113368 Sandwich ELISA Serum, Plasma, Biological Fluids 31.25-2000 pg/mL 31.25 pg/mL Log in to see
11.926027 ABIN2708069 Sandwich ELISA Cell Culture Supernatant, Serum, Plasma 15.6-1000 pg/mL 15.6 pg/mL Log in to see
1 ABIN3032886 Sandwich ELISA Cell Culture Supernatant, Plasma, Serum 5.5-4000 pg/mL 5.5 pg/mL Log in to see
1 ABIN491735 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.62-1000 pg/mL 15.62 pg/mL Log in to see
1 ABIN511226 Sandwich ELISA Serum, Cell Culture Supernatant, Tissue Homogenate, Plasma, Body Fluids 100-2500 pg/mL 100 pg/mL Log in to see
1 ABIN1115707 Sandwich ELISA Serum, Plasma, Biological Fluids 31.25-2000 pg/mL 31.25 pg/mL Log in to see
1 ABIN1568493 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.625-1000 pg/mL 15.625 pg/mL Log in to see
1 ABIN2683490 Sandwich ELISA Cell Culture Supernatant, Plasma, Serum, Urine Log in to see 1
1 ABIN2683489 Sandwich ELISA Plasma (heparin), Plasma (EDTA), Cell Culture Supernatant, Milk, Saliva, Serum 15.6-1000 pg/mL 15.6 pg/mL Log in to see
1 ABIN2541214 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 15.62-1000 pg/mL 15.62 pg/mL Log in to see
1 ABIN4967067 Sandwich ELISA Cell Culture Supernatant, Plasma, Tissue Homogenate, Serum, Cell Lysate, Biological Fluids 15.625-1000 pg/mL 15.625 pg/mL Log in to see
1 ABIN455651 31.2-2000 pg/mL 31.2 pg/mL Log in to see
1 ABIN2115923 Log in to see

Top referenced Amphiregulin ELISA Kit for ELISA

Similar Amphiregulin ELISA Kits

Application / Reactivity Human
ELISA 17 ELISA Kits

General

Antigen Amphiregulin (AREG) ELISA Kits
Reactivity Human
Kits with alternative reactivity to:
(18), (17), (12), (7), (7), (7), (7), (3), (3), (2), (1), (1), (1), (1)
Method Type Sandwich ELISA
Detection Range 10-4000 pg/mL
Minimum Detection Limit 10 pg/mL
Application ELISA
Pubmed 8 references available
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Product Details Amphiregulin ELISA Kit

Target details Application Details Handling References for Amphiregulin Kit (ABIN624943) Images
Purpose Human Amphiregulin ELISA Kit for cell culture supernatants, plasma, and serum samples.
Sample Type Plasma, Cell Culture Supernatant, Serum
Analytical Method Quantitative
Specificity This ELISA kit shows no cross-reactivity with the following 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-6, IL-7, IL-8, IL-9, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), 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.
Sensitivity < 10 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

Target details

Product Details Amphiregulin ELISA Kit Application Details Handling References for Amphiregulin Kit (ABIN624943) Images back to top
Antigen
Alternative Name AR (amphiregulin) (AREG ELISA Kit Abstract)
Background The Human Amphiregulin (AR) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human AR in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human AR coated on a 96-well plate. Standards and samples are pipetted into the wells and AR present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human AR 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 AR 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 374
UniProt P15514
Research Area Cancer, Growth Factors
Pathways RTK Signaling, EGFR Signaling Pathway

Application Details

Product Details Amphiregulin ELISA Kit Target details Handling References for Amphiregulin Kit (ABIN624943) Images back to top
Application Notes Recommended Dilution for serum and plasma samples2 fold
Sample Volume 100 μL
Plate Pre-coated,Strips (12 x 8)
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 A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B should be used for dilution of culture supernantants and urine. Suggested dilution for normal serum/plasma: 2 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 580 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture supernates/urine) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL AR standard (50 ng/mL) from the vial of Item C, into a tube with 460 µL Assay Diluent A or 1x Assay Diluent B to prepare a 4,000 pg/mL standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the 4,000 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). 40 µL standard + 460 µL 200 µL 200 µL 200 µL 200myl 200 µL 4,000 1333 444.4 148.1 49.38 16.46 0 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 500-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 20 µL of HRP-Streptavidin concentrate into a tube with 10 ml 1x Assay Diluent B to prepare a final 500 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 A Human AR concentration (pg/mL) 10 100 1000 10000 O D =4 50 n m 0.001 0.01 0.1 1 10 Assay Diluent B Human AR concentration (pg/mL) 10 100 1000 10000 O D =4 50 n m 0.001 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of AR is typically less than 10 pg/mL.
Recovery: Recovery was determined by spiking various levels of human AR into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 85.0 74-95 Plasma 72.7 64-87 Cell culture media 100.4 89-111
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 106.5 112.6 104.1 Range ( %) 95-116 101-120 82-103 1:4 Average % of Expected 112.4 115.7 108.5 Range ( %) 96-118 102-124 94-117
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

Product Details Amphiregulin ELISA Kit Target details Application Details References for Amphiregulin Kit (ABIN624943) Images back to top
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

References for Amphiregulin Kit (ABIN624943)

Product Details Amphiregulin ELISA Kit Target details Application Details Handling Images back to top
Product cited in:

Morancho, Martínez-Barriocanal, Villanueva, Arribas: "Role of ADAM17 in the non-cell autonomous effects of oncogene-induced senescence." in: Breast cancer research : BCR, Vol. 17, pp. 106, 2015

Miyabayashi, Ijichi, Mohri, Tada, Yamamoto, Asaoka, Ikenoue, Tateishi, Nakai, Isayama, Morishita, Omata, Moses, Koike: "Erlotinib prolongs survival in pancreatic cancer by blocking gemcitabine-induced MAPK signals." in: Cancer research, Vol. 73, Issue 7, pp. 2221-34, 2013

Miyabayashi, Ijichi, Mohri, Tada, Yamamoto, Asaoka, Ikenoue, Tateishi, Nakai, Isayama, Morishita, Omata, Moses, Koike: "Erlotinib prolongs survival in pancreatic cancer by blocking gemcitabine-induced MAPK signals." in: Cancer research, Vol. 73, Issue 7, pp. 2221-34, 2013

Miyabayashi, Ijichi, Mohri, Tada, Yamamoto, Asaoka, Ikenoue, Tateishi, Nakai, Isayama, Morishita, Omata, Moses, Koike: "Erlotinib prolongs survival in pancreatic cancer by blocking gemcitabine-induced MAPK signals." in: Cancer research, Vol. 73, Issue 7, pp. 2221-34, 2013

Miyabayashi, Ijichi, Mohri, Tada, Yamamoto, Asaoka, Ikenoue, Tateishi, Nakai, Isayama, Morishita, Omata, Moses, Koike: "Erlotinib prolongs survival in pancreatic cancer by blocking gemcitabine-induced MAPK signals." in: Cancer research, Vol. 73, Issue 7, pp. 2221-34, 2013

Hristozova, Konschak, Budach, Tinhofer: "A simple multicolor flow cytometry protocol for detection and molecular characterization of circulating tumor cells in epithelial cancers." in: Cytometry. Part A : the journal of the International Society for Analytical Cytology, Vol. 81, Issue 6, pp. 489-95, 2012

Hirota, Moreau, Iablokov, Dicay, Renaux, Hollenberg, MacNaughton: "Epidermal growth factor receptor transactivation is required for proteinase-activated receptor-2-induced COX-2 expression in intestinal epithelial cells." in: American journal of physiology. Gastrointestinal and liver physiology, Vol. 303, Issue 1, pp. G111-9, 2012

Oliveras-Ferraros, Cufí, Queralt, Vazquez-Martin, Martin-Castillo, de Llorens, Bosch-Barrera, Brunet, Menendez: "Cross-suppression of EGFR ligands amphiregulin and epiregulin and de-repression of FGFR3 signalling contribute to cetuximab resistance in wild-type KRAS tumour cells." in: British journal of cancer, Vol. 106, Issue 8, pp. 1406-14, 2012

Images

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