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Human EGF ProQuantum Immunoassay Kit
How ProQuantum immunoasssays 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 annea... View More
(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 annea... View More
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Product Specifications
Analytical sensitivity
<0.01 pg/mL
Assay range
0.0128 - 5000 pg/mL
Sample type/volume
Serum
2-5 µL
Supernatant
2-5 µL
Plasma
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
EGF
Species (tested)
Human
Assay kit format
Proximity-based Immunoassay
Label or dye
FAM™
About This Kit
The ProQuantum Human EGF Immunoassay Kit is designed to provide highly sensitive quantitative measurements of Human EGF protein in small sample volumes. Utilizing proximity ligation assay (PLA) 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 Cloud-based 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 below).
The assay workflow is fast and easy-2 steps in 2 hours. There are a total of 7 components in each kit (figure below). 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 cloud-based software 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, set up customized assay instructions, and obtain robust statistical group-wise comparisons.
Target information
EGF (Epidermal growth factor) exerts its actions by binding to the EGF Receptor, a 170 kDa protein kinase. Activation of EGFR initiates diverse cellular pathways in response to toxic environmental stimuli, or to EGF binding to the receptor, the EGFR forms homo- or heterodimers with other family members. Each dimeric receptor complex initiates a distinct signaling pathway by recruiting different Src homology 2 (SH2) containing effector proteins. EGF is far and wide expressed in kidney, cerebrum, prostrate and salivary glands. EGF acts as a potent mitogenic factor and the phosphorylated receptor recruits adapter proteins like GRB2 that activates complex downstream signaling cascades. EGF activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STAT modules. Research studies suggest the protein may also play important role in activating the NF-kappa-B signaling cascade. Defects in the EGGF gene are the cause of hypomagnesemia type 4 and dysregulation has been associated with the growth and progression of certain cancers.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Gene aliases : EGF, HOMG4, URG
Gene ID : (Human) 1950
Gene symbol : EGF
Protein Aliases : beta-urogastrone, EGF, Urogastrone, Pro-epidermal growth factor
UniProt ID (Human) P01133
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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