Human HGF R/c-MET DuoSet ELISA

Catalog # Availability Size / Price Qty
DY358
Ancillary Products Available
HGF R/c-MET ELISA Kit ELISA
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Product Details
Procedure
Citations (6)
FAQs
Supplemental Products
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Human HGF R/c-MET DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human HGF R/c-Met. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Data Example

Human HGF R / c-MET ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: HGF R/c-MET

HGF receptor, a product of the proto-oncogene c-met, is a heterodimeric transmembrane glycoprotein that is a receptor-type tyrosine kinase. c-MET is synthesized as a single-chain precursor (pr170) which undergoes post-translational glycosylation and proteolytic cleavage to give rise to the heterodimeric mature form.

Long Name:
Hepatocyte Growth Factor Receptor
Entrez Gene IDs:
4233 (Human); 17295 (Mouse)
Alternate Names:
AUTS9; cMET; c-MET; EC 2.7.10; EC 2.7.10.1; hepatocyte growth factor receptor; HGF R; HGF receptor; HGF/SF receptor; HGFR; Met (c-Met); met proto-oncogene (hepatocyte growth factor receptor); met proto-oncogene tyrosine kinase; MET; oncogene MET; Proto-oncogene c-Met; RCCP2; Scatter factor receptor; SF receptor; Tyrosine-protein kinase Met

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human HGF R/c-MET DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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  1. Tumor Inhibitory Effect of IRCR201, a Novel Cross-Reactive c-Met Antibody Targeting the PSI Domain
    Authors: H Park, D Kim, E Kim, JK Sa, HW Lee, S Yu, J Oh, SH Kim, Y Yoon, DH Nam
    Int J Mol Sci, 2017;18(9):.
    Species: Human
    Sample Types: Cell Lysates
  2. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016;1(13):.
    Species: Human
    Sample Types: Urine
  3. Cbl-independent degradation of Met: ways to avoid agonism of bivalent Met-targeting antibody.
    Authors: Lee, J M, Kim, B, Lee, S B, Jeong, Y, Oh, Y M, Song, Y-J, Jung, S, Choi, J, Lee, S, Cheong, K H, Kim, D U, Park, H W, Han, Y K, Kim, G W, Choi, H, Song, P H, Kim, K A
    Oncogene, 2014;33(1):34-43.
    Species: Human
    Sample Types: Cell Lysates
  4. Integrated proteomic analysis of human cancer cells and plasma from tumor bearing mice for ovarian cancer biomarker discovery.
    Authors: Pitteri SJ, JeBailey L, Faca VM, Thorpe JD, Silva MA, Ireton RC, Horton MB, Wang H, Pruitt LC, Zhang Q, Cheng KH, Urban N, Hanash SM, Dinulescu DM
    PLoS ONE, 2009;4(11):e7916.
    Species: Human
    Sample Types: Plasma
  5. Urine analysis and protein networking identify met as a marker of metastatic prostate cancer.
    Authors: Russo AL, Jedlicka K, Wernick M, McNally D, Kirk M, Sproull M, Smith S, Shankavaram U, Kaushal A, Figg WD, Dahut W, Citrin D, Bottaro DP, Albert PS, Tofilon PJ, Camphausen K
    Clin. Cancer Res., 2009;15(13):4292-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Plasma level of soluble c-Met is tightly associated with the clinical risk of preeclampsia.
    Authors: Zeng X, Sun Y, Yang HX, Li D, Li YX, Liao QP, Wang YL
    Am. J. Obstet. Gynecol., 2009;201(6):618.e1-7.
    Species: Human
    Sample Types: Plasma

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