Category: Technical Spotlight

iLite® Assay Ready Cells are developed by our partners at Svar Life Science. The cell based solutions are based on the iLite technology, a cleverly engineered cell-based assay system with a dual reporter gene readout. They offer the ease of use and robustness of a Ligand Binding Assay and can be developed for virtually any pharmaceutical target and allows an easy, rapid and accurate test format for measurement and quantification of drug activity and immunogenicity.

Key Benefits

Assay Ready Format:
iLite cells are delivered as Assay Ready Cells, and stored at -80°C. There is no need for cell culturing and continuous maintenance of cells – just thaw and dilute before use in the assay.

Normalization Gene:
A second reporter gene used as an internal control will compensate for differences in cell number. In addition, it can be used to compensate for serum matrix effects, other complex matrices or if luciferase is sensitive to the compound you are analyzing.

Sensitivity and Specificity:
The sensitivity of iLite cells is enhanced through up-regulation of key components, such as the receptor and certain signaling pathway proteins. The up-regulation of receptors also confers a higher specificity, and this is enhanced through the use of chimeric transcription factors and synthetic reporter gene promoters.

These elements are used as a lock and key to transcription of the reporter gene – only the chimeric transcription factor can bind to the synthetic promoter, and endogenous transcription factors are thereby unable to trigger expression of the reporter gene. In this way, pathway cross-talk can be effectively minimized.

Precision:
The Assay Ready Format reduces assay variability in comparison with cells in culture. Traditionally, cell-based assays are known for their high variability, with %CV often over 25%. Robustness assays of iLite assays have shown that repeatability and intermediate precision are in the range of 4-11% CV, and sample accuracy between 92-107%.

Find all of the iLite products offered on the iLite Assay Ready Cells product page.

Eagle Bioscience’s is excited to highlight the Alpha Synuclein Preformed Fibrils (Type 1) from StressMarq!

About the Alpha Synuclein Pre-Formed Fibrils (Type 1)

StressMarq’s Alpha Synuclein Pre-formed Fibrils (Type 1) are produced with low endotoxin levels, making them the suitable choice for both in-vivo and in-vitro work. Alpha Synuclein PFFs seed the formation of new fibrils from active alpha synuclein monomers, and can be used to induce endogenous alpha synuclein phosphorylation and subsequent Lewy body inclusion formation in neuronal cell culture or for in vitro oligomerization studies.

Background

Alpha-Synuclein (SNCA) is expressed predominantly in the brain, where it is concentrated in presynaptic nerve terminals¹. Alpha-synuclein is highly expressed in the mitochondria of the olfactory bulb, hippocampus, striatum and thalamus². Functionally, it has been shown to significantly interact with tubulin³, and may serve as a potential microtubule-associated protein. It has also been found to be essential for normal development of the cognitive functions; inactivation may lead to impaired spatial learning and working memory⁴. SNCA fibrillar aggregates represent the major non A-beta component of Alzheimers disease amyloid plaque, and a major component of Lewy body inclusions, and Parkinson’s disease. Parkinson’s disease (PD) is a common neurodegenerative disorder characterized by the progressive accumulation in selected neurons of protein inclusions containing alpha-synuclein and ubiquitin^{5, 6}.

ATTO633 fluorescently-labelled alpha synuclein PFFs (SPR-322) were taken up, transported into the soma, and induced alpha synuclein aggregation in mouse neurocortical primary cells. (A) Neurites filled with fluorescently-labelled alpha synuclein seeds in a microfluidic co-culture system after 24 hours. (B) Alpha synuclein seeds within the soma and neurites of mouse neurocortical primary cells after 24 hours. Experiment and imaging courtesy of Cellectricon.

References

1. “Genetics Home Reference: SNCA”. US National Library of Medicine. (2013).
2. Zhang L., et al. (2008) Brain Res. 1244: 40-52.
3. Alim M.A., et al. (2002) J Biol Chem. 277(3): 2112-2117.
4. Kokhan V.S., Afanasyeva M.A., Van’kin G. (2012) Behav. Brain. Res. 231(1): 226-230.
5. Spillantini M.G., et al. (1997) Nature. 388(6645): 839-840.
6. Mezey E., et al. (1998) Nat Med. 4(7): 755-757.

The Alpha Synuclein Preformed Fibrils (Type 1) were utilized in the following studies:

• Deciphering the role of hsp110 chaperones in diseases of protein misfolding. Yakubu, U. M. (2021) PhD Thesis, University of Texas.
• Suppression of aggregate and amyloid formation by a novel intrinsically disordered region in metazoan Hsp110 chaperones. Yakubu, U.M., Morano, K.A. (2021) J Biol Chem Jan-Jun 2021;296:100567.
• Design, Synthesis and Chemically Engineered Graphene Quantum Dot Applications: Contrast Agent for MR Imaging and Targeted Therapeutics on Parkinson’s Treatment. Poonkuzhali, K. et al. (2022) SSRN 4056733.
• Rational Generation of Monoclonal Antibodies Selective for Pathogenic Forms of Alpha-Synuclein. Gibbs, E. et al. (2022) Biomedicines 10,2168.
• LRP1 is a neuronal receptor for α-synuclein uptake and spread. Chen, K. et al. (2022) Mol Neurodegener 17(1):57.
• Granulovacuolar degeneration bodies are independently induced by tau and α-synuclein pathology. Jorge-Oliva, M. et al. (2022) Alzheimer’s Research and Therapy (14)187.

If you have any questions about this product or any of our other offerings, contact us here.

The Genetic Analysis GA-Map® Dysbiosis Text Lx has been validated on the Luminex MAGPIX system. The company proudly announced the successful completion of validation at the beginning of February. Previously, the GA-Map Dysbiosis Test Lx was only validated for the Luminex 200.

GA-Map Dysbiosis Test Lx Principle

The GA-Map Dysbiosis Test Lx is a tool that allows mapping of the intestinal microbiota profile for a selected set of bacteria in a sample. The GA-Map Dysbiosis Test is based on advances in DNA profiling using probes targeting variable regions (V3 to V7) of the bacterial 16S rRNA gene to characterize and identify bacteria present. The targets are identified in a molecular multiplex assay that utilizes the Single Nucleotide Primer Extension (SNuPE) technology patented by professor Knut Rudi of the Norwegian University of Life Sciences (​NMBU) (US6617138). A unique algorithm takes advantage of all the data generated by the detection of the SnuPE products to determine the microbiota profile in the sample. The algorithm is incorporated in the GA-Map Dysbiosis Analyzer software that accompanies the test.

Luminex MAGPIX Validation

The MAGPIX® instrument is the most affordable instrument of the Luminex’s xMAP instruments. The MAGPIX system is already a widely used platform and a large number of laboratories have already installed this system for performing other tests.

The human microbiome market is accelerating both in terms of evidence-based research and pharma products launched. The market’s need for a validated microbiome test is growing with this market. Thus, the news today that more labs can start microbiome testing with GA-Map on MAGPIX will fuel this growth and create more research opportunities.

Check out the full press release here.

If you have any questions about the GA-Map Dysbiosis Test Lx or our other offerings, contact us here.

New data suggests the Cov19 FluoBolt-DAT detects antibodies against the new Omicron variant.

Quantitative Duplex Antibody Test (DAT) for antibodies to SARS-CoV-2

The Cov19 FluoBolt-DAT allows the simultaneous and quantitative measurement of antibodies against the S1 RBD AND Nucleocapsid antigen of SARS-CoV-2. In contrast, other antibody assays detect all antibodies generated against the S1- OR NC-protein. This test, however, is epitope specific. It detects immune-dominant antibodies species, i.e. the most important antibodies generated by either vaccination or infection.

This is demonstrated by the fact, that it detects anti-S1RBD against all dominant virus variants identified so far. The data indicates (see chart below) that the S1RBD part of the Cov19 FluoBolt-DAT assay detects the immune-dominant epitope and that individuals testing S1RBD -positive in the FluoBolt-DAT test have developed significant amounts of antibodies against this epitope. Therefore, they might be well protected against all current and future SARS-CoV-2 variants

Benefits of the Cov19 FluoBolt-Dat Assay:

• • Only One Single Measurement Required
  • Small Sample Size Need – 10 uL
  • Results within 60 minutes

  Other Cov19 FluoBolt-DAT Resources:

  • Quick Guide
  • New Product Announcement
  • Literature

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  Check out our other COVID-19 assays here.

  If you have any questions about this kit or our other offerings, contact us here.

This week Eagle Biosciences is highlighting a revolutionary test in the microbiome industry. The GA-map® Dysbiosis Test Lx was developed by Genetic Analysis in Norway. The GA-map ® is a tool that allows mapping of the intestinal microbiota profile for a selected set of bacteria in a sample. The test is based on advances in DNA profiling using probes targeting variable regions (V3 to V7) of the bacterial 16S rRNA gene to characterize and identify bacteria present. The targets are identified in a molecular multiplex assay that utilizes the Single Nucleotide Primer Extension (SNuPE) technology patented by professor Knut Rudi of the Norwegian University of Life Sciences (​NMBU) (US6617138). A unique algorithm takes advantage of all the data generated by the detection of the SnuPE products to determine the microbiota profile in the sample. The algorithm is incorporated in the GA-map® Dysbiosis Analyzer software that accompanies the test.

Check out this video for a quick overview of the GA-map ® Dysbiosis Test Lx Platform equipment and techniques.

If you have any questions about the GA-map® Dysbiosis Test Lx or our other offerings contact us here.