Author: Eagle Biosciences

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Folic Acid ELISA Highlighted in Recent Study!

by Eagle Biosciences

The Eagle Bioscience’s Folic Acid ELISA Assay Kit Highlighted in a recent study! The study was conducted to assess the disruption in nutritional attributes of Capsicum annuum by five different microplastic (MP) types. Check out the abstract and full text below.

Abstract

This study was executed to assess the disruption in nutritional attributes of Capsicum annuum by five different microplastic (MP) types i.e. polyvinylchloride (PVC), polystyrene (PS), high-density polyethylene (HDPE), low-density polyethylene (LDPE), and polyethylene terephthalate (PET). Significant (P ≤ 0.05) differences were recorded among traits of C. annuum fruit concerning the type of MPs. PVC was more hazardous than the other MPs and depleted maximum protein content. Likewise, HDPE exerted a 51.62% reduction in carbohydrates compared to the control. Vitamin-A (-32.09%) and vitamin-B6 (-37%) were severely influenced by the PVC in C. annuum fruit. HDPE and PVC both declined the oleic acid contents. PVC caused a 47.6% reduction in linoleic acid while the least damaging was the PET (4.71%) for this attribute. Palmitic acid and stearic acid were however more negatively affected by the HDPE. The PVC and HDPE severely degraded the total flavonoid contents and phenolics. Macro and micronutrients of C. annuum fruit were also negatively influenced by all the five MPs used in this study. HDPE, PVC and LDPE, respectively caused significant repression in Ca, K, Mg and Zn (P ≤ 0.05). Overall, HDPE and PVC caused significant damage and decreased nutritional contents. This is, so far, the premier study unraveling the changes in the nutrition of C. annuum fruits due to MPs. Accordingly, further research is highly recommended on the changes caused by MPs in the nutritional value of food crops.

Khadiga Alharbi, Muhammad Aqeel, Noreen Khalid, Atia Nazir, Muhammad Kashif Irshad, Fahad Mohammed Alzuaibr, Haifa AbdulAziz Sakit AlHaithloul, Noreen Akhter, Omar Mahmoud Al-Zoubi, Muhammad Qasim, Khalid M.Al Syaad, Manal Abdullah AlShaqhaa, Ali Noman. Microplastics in soil differentially interfere with nutritional aspects of chilli peppers. South African Journal of Botany. Volume 160. 2023. Pages 402-413. ISSN 0254-6299, https://doi.org/10.1016/j.sajb.2023.07.027.

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Microscopy Product Spotlight: SpheroTribe

by Eagle Biosciences

What is it SpheroTribe intended for?

SpheroTribe provides a simple toolkit to generate consistent and robust 3D cell structures. Simply dilute the SpheroTribe solution into your culture medium of choice, watch your cells turn into uniformly sized 3D spheroids and collect them for your downstream assays.

Once diluted in your culture medium of choice, our concentrated polymer-based solution increases the medium viscosity favoring cell-cell contacts. SpheroTribe offers a simple method to generate homogeneous 3D cell structures with increased control over their size and shape, which can be easily handled and washed for downstream experiments.

SpheroTribe is particularly useful to boost aggregation when working with challenging cells, minimize variability between samples and improve the consistency of your migration/invasion assays, immunostaining, drug screening or in vivo implantation experiments.

SpheroTribe improves cardiac organoid formation by boosting hiPSC aggregation

U-87 glioblastoma cells were seeded at 1,000 cells per well in round-bottom wells molded in agarose using a Stampwell U-shape (Idylle) in full culture medium without (right) or added with SpheroTribe (left). After 3 days, pictures were taken and number of cell aggregates per well were quantified from 64 independent wells and associated standard deviation (SD) values were calculated.

Kit Description:

You can purchase just the methylcellulose solution or choose among 2 different size kits:

25mL kit contents:

  • 25mL of 5X methylcellulose solution
  • 10x U-bottom 96-well plates
  • 2x racks of 96 pipette tips (200µL) with a large opening
 2.5mL kit contents:

    • 2.5mL of 5X methylcellulose solution
    • 1x U-bottom 96-well plate
    • 20 pipette tips (200µL) with a large opening

Applications

SpheroTribe has been successfully used for spheroid/organoid formation with the following cell types:
Patient-derived stem-like glioblastoma cells (GB P3 and BL13), human glioblastoma cell lines (U87 & T98G), HeLa, human vaginal mucosal melanoma (HMV-II), human primary colorectal cancer cells, human breast cancer cells (MDA-MB 231), human induced pluripotent stem cells, monkey kidney fibroblast-like cell line (COS-7), primary neurons from rat embryos (E18) & murine melanoma cells (B16F10).

Experimental assays:
Once spheroids have grown to your desired size, you can use them for any kind of assay according to your regular workflow. The SpheroTribe solution can be readily washed off, leaving a spheroid available for other tests at any stage of your protocol.

Immune infiltration of B16F10 spheroids after immune checkpoint blockade

A. 10,000 B16F10 cells were grown for 6 days as spheroids using SpheroTribe. B. 100,000 PBMC from murine spleen were activated with IL-15 (40 ng/mL) [1], incubated with anti-PD1 (10 µg/mL) for 1h and added on B16F10 spheroids for 3 days.
Graph shows flow cytometry quantification of differential lymphocyte infiltration after spheroid dissociation according to treatment. N=4. Mann-Whitney U Test, p-value<0.05. [1] https://doi.org/10.3389/fonc.2022.898732.

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Dopamine ELISA Utilized in Recent Publication

by Eagle Biosciences

The Eagle Bioscience’s Dopamine ELISA Assay Kit was utilized in a recent publications! The aim of the study was to investigate the potential of iron oxide nanoparticles produced using ascorbic acid (AA-IONPs) against Parkinson’s Disease. Check out the abstract and full text below!

Abstract

One of the most prevalent neurological movement diseases affecting the geriatric population globally is Parkinson’s disease (PD). Recent studies have highlighted the potency of biomolecules in the generation of nanomaterials and also over their impact on neuroprotection. The objective of this research was to investigate the potential of iron oxide nanoparticles produced using ascorbic acid (AA-IONPs) against PD. Numerous analytical methods including UV–Vis analysis, Fourier-Transform Infrared Spectroscopy (FTIR), dynamic light scattering (DLS), and electron microscopy (SEM, TEM), were used to analyze the produced AA-IONPs. Nitric oxide, prostaglandin E2, and inflammatory cytokines analyses such as IL-6 and IL-1 were employed to assess the neuroprotective effect of synthesized AA-IONPs on inflammatory agent lipopolysaccharides driven murine microglial BV2 cells. And also Parkinson-induced C57BL/6 mice were given the nanoparticle treatment to confirm the in vivo effects of the produced nanoparticles. Our characterization findings had demonstrated that AA-IONPs have a significant role in acting as an ideal nano drug and may have the ability to reduce inflammation in in vitro murine microglial BV2. The outcomes of in vivo tests conclusively show that AA-IONPs had reduced neuroinflammation and enhanced motor coordination in Parkinson’s disease-induced rats.

Li, L., Luo, P., Wu, S. et al. Deciphering the neuroprotective effect of ascorbic acid mediated synthesis of iron oxide nanoparticles against Parkinson’s disease: an in vitro and in vivo approach. Macromol. Res. 31, 949–960 (2023). https://doi.org/10.1007/s13233-023-00186-x

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Human IL-6 High Sensitive ELISA Utilized in a Recent Publication

by Eagle Biosciences


The Biomedica Human IL-6 High Sensitive ELISA Assay Kit was highlighted in a recent publication that explored factors associated with incident vertebral fractures in glucocorticoid-treated Duchenne muscular dystrophy. Check out the abstract and full text below!

Abstract

Purpose: Prevention of fractures is an unmet need in glucocorticoid (GC)-treated Duchenne muscular dystrophy. This study explored factors associated with incident vertebral fractures (VFs) to inform future fracture prevention efforts.

Methods: VFs were evaluated prospectively at study baseline and 12 months on lateral spine radiographs in participants aged 4 to 25 years with Duchenne muscular dystrophy. Clinical factors were analyzed for their association with the change in Spinal Deformity Index (sum of the Genant-defined VF grades from T4 to L4) between baseline and 12 months.

Results: Thirty-eight males were evaluated (mean ± SD age at baseline 11.0 ± 3.6 years; mean ± SD GC duration at baseline 4.1 ± 3.1 years; 74% ambulatory). Nine of 38 participants (24%) had 17 incident VFs, of which 3/17 VFs (18%) were moderate/severe. Participants with 12-month incident VF had lower mean ± SD baseline lumbar spine areal bone mineral density Z-scores (-2.9 ± 1.0 vs -1.9 ± 1.1; P = .049) and lower total body less head areal bone mineral density Z-scores (-3.1 ± 1.2 vs -1.6 ± 1.7; P = .036). Multivariable linear regression showed that at least 1 VF at baseline (P < .001), a higher number of antecedent non-VF (P < .001), and greater bone age delay at baseline (P = .027) were significant predictors of an increase in the Spinal Deformity Index from baseline to 12 months.

Conclusion: The observation that ≥ 1 prevalent VF and/or non-VF were the strongest predictors of incident VFs at 12 months supports the need for prevention of first fractures in this high-risk setting. Bone age delay, a marker of GC exposure, may assist in the prioritization of patients in efforts to prevent first fractures.

Keywords: Duchenne muscular dystrophy; bone fragility; glucocorticoids; incident fractures; osteoporosis; vertebral fractures.

Risk Factors Associated with Incident Vertebral Fractures in Steroid-treated Males with Duchenne Muscular Dystrophy . Phung K, McAdam L, Ma J, McMillan HJ, Jackowski S, Scharke M, Matzinger MA, Shenouda N, Koujok K, Jaremko JL, Wilson N, Walker S, Hartigan C, Khan N, Page M, Robinson ME, Saleh DS, Smit K, Rauch F, Siminoski K, Ward LMJ Clin Endocrinol Metab. 2023 Aug 23:dgad435. doi: 10.1210/clinem/dgad435. Epub ahead of print.PMID: 37610420

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Microscopy Product Spotlight: AgarSqueezer

by Eagle Biosciences

What is it AgarSqueezer intended for?

AgarSqueezer is a microscope slide chamber equipped with a molded agar-based compression system. Use it to assess cell response to short and long-term mechanical confinement within a physiological rigidity range.

It is very helpful if you want to analyze how your cells will react if you squeeze them for a prolonged period. Or if you want to study how mechanical confinement affects drug cell resistance. And if you want to perform immunostaining in situ.

Kit Description:

You can choose among 2 kits:

  • The “1 AgarSqueezer kit” contains 1 AgarSqueezer device
  • The “2 AgarSqueezers kit” contains 2 AgarSqueezers devices.

In addition to the device, each kit contains also:

  • 1 x insert to hold up to 2 AgarSqueezers on the microscope stage
  • 1 x 16G flat cut needle to make holes in the agar gel, facilitating diffusion of culture medium or drugs during the experiment
  • 1 x 20G flat cut needle (same)

For users who also need a wafer to mold agarose, contact us to learn about the four different heights we offer.

Features

  1. Tunable stiffness in a physiological range [1-150] kPa – Use of agarose as a cheap and biocompatible polymer.
  2. Open access to the reservoir – Possibility to add drugs, and reagents. Easy medium renewal.
  3. Autoclavable and reusable systems.
  4. Compatibility with multiple microscopy techniques – Confocal, spinning, super-resolution. Open access for microscope objectives. Use of optical glass coverslip to make cells grow.
  5. Easy to recover coverslip with cells for subsequent molecular analysis – FACS, qPCR, Western-Blot, Immunoflourescence (possible in situ).
  6. Long-term analysis of the cell adaptation to confinement – Up to several days, for time-lapse studies.
  7. Study of the specific impact of mechanical loads on the biology of cells – Gas permeability of the system allows to get rid of the hypoxia conditions.
  8. Easy to assemble and disassemble the system

For more information about this product or any others from the Microscopy line, contact us here.

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Eagle Biosciences will be at ASN Kidney Week in Philadelphia, PA!

by Eagle Biosciences

Eagle Biosciences will be at ASN Kidney Week in Philadelphia, PA!

Eagle Biosciences will be at ASN Kidney Week next week, Thursday, November 2 to Sunday, November 5, at the Pennsylvania Convention Center. Stop by booth #1642 to learn more about Intact FGF23, Fetuin A, and other assays that could help you with your kidney related research! We’ll be there to answer any questions you may have, or stop and say hi! We love seeing our customers!

Product Highlights

Intact FGF23: This CLEIA ELISA is a unique and highly accurate assay for the measurement of FGF23, which has been linked to impaired renal function.

Fetuin A (PTM): This ELISA is a unique immunoassay for the measurement of Fetuin-A with specific post translational modifications in urine samples. This biomarker has shown to be an early indicator of diabetic kidney disease (DKD).

If you have questions about either of these products or any of our other offerings, contact us here.

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New Products for Host Cell Protein Detection

by Eagle Biosciences

Eagle Biosciences is excited to announce the newest product series for Host Cell Protein Detection!

Host cell proteins (HCPs) are a major class of impurities produced during biotherapeutic manufacturing. They must be removed from the final drug product to both assure patient safety and maintain drug efficacy. Our wide range of Host Cell Protein Detection Kits are easy to use and highly sensitive.

What are HCPs and why must they be removed from biologic drugs?

HCPs are proteins produced or encoded by the host organisms used to produce recombinant therapeutic proteins. Genetic engineering allows the host organism cells to be transformed to produce a protein of interest. During the recombinant protein production, host cells also coproduce proteins related to the normal cell functions such structural proteins, as well as proteins required for normal cellular growth and function, and vary in both number and concentration depending on the chosen host species and the manufacturing process being used. In general, apart from the therapeutic protein of interest, all endogenous proteins co-expressed by the host cells are called host-cell proteins.

Why must HCPs be removed from biologic drugs?

HCPs must be removed from the final biotherapeutic product to avoid adverse effects. Almost all HCPs carry safety risks as foreign proteins due to the potential to elicit immune response in humans (e.g, cytokine storm). In addition, some HCPs can also act to enhance the immune response to a drug product. Certain HCPs can also affect drug product stability and efficacy if not adequately removed or inactivated.

How are HCPs detected?

ELISAs are widely used for detecting HCPs, where they are generally configured in a sandwich assay format for improved specificity. In this scenario, a microplate-bound antibody is used for analyte capture, then a second analyte-specific antibody (that binds a different epitope on the target molecule) is added to enable detection. By incorporating a reference standard (e.g., a purified protein) into the assay design, it is possible to quantify the analyte of interest and confirm that its concentration meets regulatory requirements. Advantages of ELISA are that it is sensitive and compatible with high sample throughput – key considerations for biopharmaceutical manufacturing.

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

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More Complement Assays!

by Eagle Biosciences

Eagle Biosciences is excited to bring you a wide array of more complement assays!

The complement system is an essential part of the immune system in the human body, playing a crucial role in defending against infections, clearing cellular debris, and promoting inflammation. It consists of a complex network of proteins and molecules that work together to enhance the immune response. The complement system can be activated through three main pathways, the classical pathway, the alternative pathway, the lectin pathway. The complement system is tightly regulated to prevent excessive immune responses and potential damage to host tissues. Various regulatory proteins are involved in controlling the activation and amplification of the complement cascade to maintain a delicate balance between defense and protection.

New Complement Assays

Rat Classical Complement Pathway ELISA Assay Kit
Rat Lectin Complement Pathway ELISA Assay Kit
Rat Alternative Complement Pathway ELISA Assay Kit
Mouse Classical Complement Pathway ELISA Assay Kit
Mouse Alternative Complement Pathway ELISA Assay Kit
Pig Classical Complement Pathway ELISA Assay Kit
Pig Lectin Complement Pathway ELISA Assay Kit
Pig Alternative Complement Pathway ELISA Assay Kit
Pig Complement Pathway ELISA Assay Kits
MASP1/C1-INH Complex ELISA Assay Kit
C3d ELISA Assay Kit
MASP-2 ELISA Assay Kit
Complement Factor H ELISA Assay Kit
Complement Factor D ELISA Assay Kit
Complement Factor I ELISA Assay Kit
Collectin-10 ELISA Assay Kit
Factor B ELISA Assay Kit
sCD59 ELISA Assay Kit
C1s/C1-INH Complex ELISA Assay Kit
Rat Terminal Complement Complex (TCC) ELISA Assay Kit
Mouse C3 ELISA Assay Kit
Mouse C1q ELISA Assay Kit
Mouse C3b ELISA Assay Kit
Mouse C4 ELISA Assay Kit

Find the complete complement product catalog here. If you have any questions about any of these products or our other offerings, contact us here.

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Eagle Biosciences will be at AACC in Anaheim!

by Eagle Biosciences

Eagle Biosciences will be at AACC in Anaheim, California!

This year AACC is in Anaheim at the Anaheim Convention Center, Sunday July 23th – Thursday July 27th. We will be at booth #1147 from July 25th to July 27th! Come by to learn about the GA-Map Dybiosis Test Lx and more assays that could help you with your microbiome or other research! We will be there to answer any questions you may have, or just stop by and say hi! We love seeing our customers!

Product Highlights

GA-Map Dysbiosis Test Lx: The first and only standardized solution for microbiome profiling! The GA-Map Dysbiosis Test Lx is a simple multiplex stool assay that maps the intestinal microbiota profile for a selected set of bacteria. The GA-map® platform uses probes that target 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 (US6617138). A unique algorithm takes advantage of all the data generated by the detection of the SnuPE products to determine dysbiosis level in the sample. The algorithm is incorporated in the GA-map® Dysbiosis Analyzer software that accompanies the test.

GA-map® Dysbiosis Test Lx Procedure Quick Guide

GA-map Dysbiosis Test

If you have any other questions about these products or our other offerings, contact us here.

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Technical Spotlight: Alpha Synuclein Preformed Fibrils (Type 1)

by Eagle Biosciences


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 ubiquitin5, 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:

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