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Human cMET ProQuantum Immunoassay Kit
ow ProQuantum immunoassays work. (A) Antibody-antigen binding: Antibodies bind to two separate epitopes on the antigen (during a 1-hr incubation), which brings the two conjugated oligonucleotides into close proximity. (B) Ligation and amplification of signal (in a qPCR instrument): DNA ligase and a third splint oligonucleotide connector are added to ligate the two ends of the conjugated oligonucleotides, creating a 100-base DNA template. Once the ligase is inactivated at 95°C, the sample is amplified through 40 cycles of annealing and exte... View More
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Product Specifications
Analytical sensitivity
<0.3 pg/mL
Assay range
0.32-25,000 pg/mL
Sample type/volume
Serum
2-5 µL
Plasma
2-5 µL
Supernatant
2-5 µL
Hands-on time
30 min
Time-to-result
2 hr
Homogenous (no wash)
Yes
Instrument
qPCR Instruments
Product size
96 Tests
Contents
Assay Dilution Buffer
Antibody-Conjugate A
Antibody-Conjugate B
Antibody-Conjugate Dilution Buffer
Standard
Master Mix
Ligase
Antibody-Conjugate A
Antibody-Conjugate B
Antibody-Conjugate Dilution Buffer
Standard
Master Mix
Ligase
Shipping conditions
Dry ice
Storage
-20°C
Protein name
c-Met
Species (tested)
Human
Assay kit format
Proximity-based Immunoassay
Label or dye
FAM™
About This Kit
The ProQuantum Human cMET Immunoassay Kit is designed to provide quantitative measurements of human cMET in small sample volumes. Utilizing proximity-based amplification technology, the assay combines the analyte specificity of high-affinity antibody-antigen binding with the signal detection and amplification capabilities of real-time PCR to achieve a simple yet powerful next-generation protein quantitation platform. A user-friendly workflow combined with intuitive software for analytics enables sample-to-answer in just 2 hours.
• High sensitivity-detect low levels of protein with greater sensitivity than traditional methods
• Broad dynamic range-5 logarithmic units, minimizing sample dilutions to ensure they fall within the range
• Small sample consumption-use 2-5 µL of sample (compared to 75 µL for triplicate wells with other methods)
• Fast, easy workflow-2 hours from sample to answer, with no wash steps
• No proprietary instrument to purchase-runs on any real-time PCR instrument
ProQuantum immunoassays utilize a matched pair of target-specific antibodies, each conjugated to a DNA oligonucleotide. During antibody-analyte binding, the two DNA oligos are brought into close proximity, which allows for ligation of the two strands and subsequent creation of a template strand for amplification. This platform leverages the sensitivity and large dynamic range of Applied Biosystems TaqMan real-time PCR technology (Figure 1).
The assay workflow is fast and easy-2 steps in 2 hours. There are a total of 7 components in each kit (Figure 2). First, mix the antibody-conjugates, dilute the sample, and create the standard curve in a working plate. Then, using a multi-channel pipette, add the antibody-conjugates and sample (or standard) into the wells of a PCR plate and incubate for 1 hour. Combine the master mix and ligase and add to the wells of the PCR plate, then run the plate on any qPCR instrument. After the run is complete, import the results file into the ProQuantum software (cloud-based version at https://apps.thermofisher.com/apps/proquantum). Using this software, the data can be analyzed easily to obtain protein concentration values. The software allows you to set up standard curves, design plate layouts, apply a 5PL weighted algorithm, and obtain robust statistical group-wise comparisons.
Target information
MET (cMET) is a receptor-like tyrosine kinase whose dysregulation has been linked to many types of human malignancies. After activation of the ligand, MET interacts with the PI3-kinase subunit PIK3R1, PLCG1, SRC, GRB2, and STAT3. There interactions lead to the activation of signaling cascades including RAS-ERK, PI3, kinase-AKT, and PLCgamma-PKC. MET plays a role in embryonic development including gastrulation, development of muscles and neurons, angiogenesis, and kidney formation. It also plays a role in adults including wound healing, organ regeneration, and tissue remodeling. MET has been linked to cancers including gastric, renal, and breast; therefore, making it a target for cancer therapeutics and diagnostic testing.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Gene aliases : AUTS9, c-Met, DFNB97, HGFR, MET, RCCP2
Gene ID : (Human) 4233
Gene symbol : MET
Protein Aliases : Hepatocyte growth factor receptor, HGF receptor, HGF/SF receptor, Proto-oncogene c-Met, Scatter factor receptor, SF receptor, Tyrosine-protein kinase Met
UniProt ID (Human) P08581
Performance Guarantee
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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