The ProQuantum Human GM-CSF Immunoassay Kit is designed to provide highly sensitive quantitative measurements of human GM-CSF 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.
GM-CSF (Granulocyte-Macrophage colony-stimulating factor) is a 14.6kDa hematopoietic growth factor that exists in glycosylated and non-glycosylated biologically active forms, and stimulates the development of granulocytes, macrophages, early megakaryocytes and eosinophil progenitor cells. The active form of the GM-CSF protein is found extracellularly as a homodimer and the GM-CSF gene has been localized to a cluster of related genes at chromosome region 5q31, which is known to be associated with interstitial deletions in the 5q-syndrome and acute myelogenous leukemia. Other genes in the cluster include those encoding interleukins 4, 5, and 13. The ability of recombinant GM-CSF to increase hematopoietic cell recovery has become a focus area in the therapeutic treatment of patients following bone marrow transplantation. Recent studies have investigated GM-CSF in inflammatory and autoimmune diseases such as arthritis, arthritis related interstitial lung disease, nephritis, and psoriasis. In the CNS, GM-CSF depletion or neutralization has been studied in experimental autoimmune encephalomyelitis (EAE) and multiple sclerosis (MS).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.