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Human NF-L ProQuantum Immunoassay Kit
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Human NF-L ProQuantum Immunoassay Kit
How 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 ext... View More
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Product Specifications
Analytical sensitivity
0.156 pg/mL
Assay range
0.48-7500 pg/mL
Sample type/volume
Plasma
2-5 µL
Serum
2-5 µL
Cerebrospinal Fluid
2-5 µL
Hands-on time
30 min
Time-to-result
2 hr
Homogenous (no wash)
No
Instrument
qPCR Instruments
Product size
5 x 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
Storage
-20°C
Protein name
NF-L
Species (tested)
Human, Mouse, Rat
Assay kit format
Proximity-based Immunoassay
Label or dye
FAM™
About This Kit
The ProQuantum NFL Immunoassay Kit is designed to provide quantitative measurements of human neurofilament light 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 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 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 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
Involved in the maintenance of neuronal caliber, neurofilaments are the intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H. Like most other intermediate filament proteins (IFPs), the expression of the different neuronal IFPs is both tissue-specific and developmentally regulated. NF-L is the light or low molecular weight microfilament subunit and runs on SDS-PAGE gels at approximately 70 kDa. Neurofilament are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H. NF-H is the heavy or high molecular weight microfilament subunit and runs on SDS-PAGE gels in the range 180-220 kDa, with some variation in different species.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Gene aliases : AI847934, CMT1F, CMT2E, NEFL, NF-L, NF68, NFL, PPP1R110
Gene ID : (Human) 4747, (Mouse) 18039, (Rat) 83613
Gene symbol : NEFL, Nefl, Nefl
Protein Aliases : 68 kDa neurofilament protein, FLJ53642, NEF3, NEFL, neurofilament, neurofilament 3, neurofilament-l, neurofilament-light, N-FL, NFM, NF-M, NLC, OTTHUMP00000196614, light molecular weight neurofilament protein, Neurofilament light polypeptide, neurofilament protein L, neurofilament protein, light chain, neurofilament subunit NF-L, Neurofilament triplet L protein, light polypeptide 68kDa, NF-L, protein phosphatase 1, regulatory subunit 110
UniProt ID (Human) P07196, (Mouse) P08551, (Rat) P19527
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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