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How does it work?

GlowMito provides a non-toxic solution to quickly visualize the entire mitochondrial network in live samples.

GlowMito quickly penetrates cells and produces a bright & stable red labeling of mitochondria without inducing cell toxicity or altering mitochondrial functions. produces a bright, red labeling of the entire mitochondrial network, including those with reduced potential. It is perfectly suitable for:

  • Live imaging: study of mitochondrial dynamics (velocity, localization), structural changes, multiplexing (potentiometric dyes, calcium signaling probes, etc)
  • Downstream analysis: flow cytometry, oxygraphy, etc

We do not recommend its use to measure mitochondrial mass or volume density.


Simplified Protocol


Frequently Asked Questions

How was mitochondrial specificity of GlowMito verified?
The strong ability of GlowMito to specifically target mitochondria has been meticulously checked by co-localization studies. More generally, the ability of lipophilic cations to specifically target mitochondria is already well-established and conjugating molecules to lipophilic cations is a commonly used method to develop mitochondria-targeted compounds.

Which assays can GlowMito be used for?
GlowMito produces a bright, red labeling of the entire mitochondrial network, including those with reduced potential. It is perfectly suitable for:

  • Live imaging: study of mitochondrial dynamics (velocity, localization), structural changes, multiplexing (potentiometric dyes, calcium signaling probes, etc)
  • Downstream analysis: flow cytometry, oxygraphy, etc

We do not recommend its use to measure mitochondrial mass or volume density.

Which types of samples has GlowMito been used with?
So far, GlowMito has been successfully used to label mitochondria in the following biological samples:

  • Human cells: HEK293, HeLa, MCF-7, MDA-MB-231, HMLE, UACC-62, U2-OS, Gli36, HAEC, SH-SY5Y, A-172, A549, patient-derived skeletal muscle cells & primary smooth muscle cells
  • Monkey cells: COS-7
  • Mice cells: primary cortical neurons
  • Tissues: hiPSC-derived heart tissues & mice isolated pressurized blood vessels
  • Parasitic protists: Trichomonas Vaginalis

GlowMito showed no internalization in yeast cells


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

The Eagle Bioscience’s NSE ELISA Assay Kit was utilized in a recent publication! This study investigated the challenges of using electrochemical immunosensors for the early detection of small cell lung cancer.


Abstract

Early and rapid detection of neuron-specific enolase (NSE) is highly significant, as it is putative biomarker for small-cell lung cancer as well as COVID-19. Electrochemical techniques have attracted substantial attention for the early detection of cancer biomarkers due to the important properties of simplicity, high sensitivity, specificity, low cost, and point-of-care detection. This work reviews the clinically relevant labeled and label-free electrochemical immunosensors developed so far for the analysis of NSE. The prevailing role of nanostructured materials as electrode matrices is thoroughly discussed. Subsequently, the key performances of various immunoassays are critically evaluated in terms of limit of detection, linear ranges, and incubation time for clinical translation. Electrochemical techniques coupled with screen-printed electrodes developing market level commercialization of NSE sensors is also discussed. Finally, the review concludes with the current challenges associated with available methods and provides a future outlook toward commercialization opportunities for easy detection of NSE.

Daisy Mehta, Divyani Gupta, Alankar Kafle, Sukhjot Kaur, and Tharamani C. Nagaiah. Advances and Challenges in Nanomaterial-Based Electrochemical Immunosensors for Small Cell Lung Cancer Biomarker Neuron-Specific Enolase. ACS Omega 2024 9 (1), 33-51 DOI: 10.1021/acsomega.3c06388


If you have any questions about the NSE ELISA Assay Kit or any of other offerings, contact us here.

What is it intended for?

DNAbsolute is composed of a ionic liquid that has a selective and high affinity for DNA strands. When mixed with your sample, the DNAbsolute reagent will directly bind DNA and precipitate it without the need for RNase treatment, protein precipitation or use of hazardous reagents.

DNAbsolute has been successfully used for extracting DNA from insects (drosophila, psyllids, beetles, ants, cochineals), plants (dried leaves) and bacteria (Salmonella). Once your sample is lysed, the whole DNA extraction process should take no more than 30 minutes.


Technical information

Bacterial species compatibility : Gram +
Tested and validated on : Staphylococcus aureus, Bacillus subtilis, Streptococcus pneumoniae.
Colour: Red
Form: liquid
Detection methods: visual & fluorescence. Also compatible with single-cell approaches (live imaging and flow cytometry)
Peak excitation/emission wavelengths: Ex:530nm/Em:650nm
Storage: at 4°C protected from light for 8 months
1 kit contains 1mL of an aqueous solution with a concentration of 1 mg/mL for a final volume of 1L of screening medium


Features of DNAbsolute

  1. Safe & Simple – No need for any columns or harmful chemicals, highly amenable to automated workflows & on-field applications
  2. Versatile – Extract DNA from a variety of samples, including some of the most challenging ones (i.e. insects, plants & corals)
  3. Extreme Sensitivity – Efficient DNA recovery from reduced starting materials, including individual specimens and even down to a single leg!

Simplified Protocol


Frequently Asked Questions

What yield can I expect?
The resulting yield can vary widely depending on the sample type and preparation. As a general rule, you can expect to recover on average 85% of DNA from a solution ranging from 10 to 100 μg/mL of DNA, and 60% of a solution ranging from 5 to 10 ng/mL of DNA. Poor quality, and/or fragmented starting material will result in reduced yield of purified DNA.

How pure will the extracted DNA be?
DNAbsolute DNA extraction method can yield highly pure DNA due to its ability to efficiently and specifically solubilize DNA, minimizing contamination with proteins, RNA, and other cellular debris. The purity of drosophila DNA extracted using DNAbsolute has been assessed by spectroscopic measurements. The average 260/280 ratio was found between 1.8-1.9 and the 260/230 ratio at 2.3.

The type of biological sample being processed can affect the extracted DNA purity. Sample lysis optimization, or additional purification steps, might be implemented when working with samples containing high levels of secondary metabolites.


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

Eagle Bioscience’s Human Ceruloplasmin ELISA Assay Kit was utilized in a recent study! The study sought to learn about the impact of sacubitril/valsartan on cardiac and systemic hypoxia in those with chronic heart failure. Check out the abstract and full text below!


Abstract

In heart failure patients with reduced ejection fraction, Sacubitril/valsartan(S/V) increased proBNPT71 glycosylation, which is regulated negatively by hypoxia via miR-30a in-vitro. Using a cohort of 73 HFrEF patients who were transitioned from standard HF medication to S/V, we found that the increase in proBNP T71 glycosylation after S/V was associated with a decrease in cardiac hypoxia. We further found that plasma levels of K709-acteylatedHIF1a, HIF-regulated and HIF-independent biomarkers also evolved consistently with a decrease in hypoxia. We further confirmed that biomarker changes were related to hypoxia, in a rat model subjected to isobaric hypoxia. We measured them in rats subjected to isobaric hypoxia. Overall, these data strongly suggest that optimally treated HFrEF patients exhibited sub clinical hypoxia that is improved by S/V. The data also posit proBNP T71 glycosylation as a biomarker of cardiac hypoxia.

Nougué, Picard, Cohen-Solal et al. Impact of sacubitril/valsartan on cardiac and systemic hypoxia in chronic heart failure. iScience (2024) 27 (1), 108520 DOI: 10.1016/j.isci.2023.108520


If you have any questions about the Human Ceruloplasmin ELISA Assay Kit or any of our other offerings, contact us here.

How does it work?

ColorFlux is a cationic compound that quickly penetrates Gram + bacteria through passive diffusion and is efficiently expelled out of the cell depending on efflux activity. ColorFlux staining was shown to reflect the activity of well-characterized efflux pumps from the major facilitator superfamily and ATP-binding cassette families in a variety of Gram+ bacteria:

  • NorA, MepA, MepB, PatA, PatB (Staphylococcus aureus & Streptococcus pneumoniae)
  • BmrA (Bacillus Subtilis)

After incubating bacteria with ColorFlux for 15 minutes, a steady-state level is reached and relative levels of efflux activity can be quickly assessed by looking at cell pellet color. It is also possible to read fluorescent signals for a precise quantification of efflux levels. Alternatively, kinetics of ColorFlux accumulation can be monitored by flow cytometry.


ColorFlux staining of MepA&B mutants in Staphylococcus aureus

Left graph shows the fluorescence signals detected for each strain using an Ex=530nm, and right graph shows corresponding fluorescence intensity values obtained for each strain at Em=645nm. WT = Wild Type strains, MepA1B+++ = MepA&B overexpressing strains.
Credits: Mrunal Patil & Jean-Michel Bolla, Aix-Marseille Université – 2023


Simplified Protocol


Frequently Asked Questions

Is ColorFlux safe to handle?
Contrary to ethidium bromide, ColorFlux is a non-toxic compound that you can safely handle on the bench and dispose of. It is a particularly useful tool for training purposes & on-field applications.

Can I use ColorFlux in Gram- bacteria?
ColorFlux has been initially developed for Gram+ bacteria because the resulting bacterial coloration after incubation with ColorFlux reflects relative levels of efflux pump activity. Indeed, the fast influx/slow efflux properties of Gram+ bacteria allow ColorFlux to be quickly internalized and then expelled as a function of efflux, resulting in different shades of red that are representative of relative levels of efflux activity.

In Gram- bacteria, the situation is a bit different: because they tend to have a slow influx/fast efflux profile, ColorFlux will be instantaneously expelled from the bacteria as soon as some pumps are present. Therefore, although bacteria without pumps will turn red, it won’t always be possible to distinguish between WT strains and those with enhanced efflux as they will always turn white. This was tested in E.coli, in which WT strain turned white and mutants with ArcAB pump deleted turned red. Some preliminary tests combining ColorFlux with compounds to reduce efflux pump activity, i.e. CCCP, have shown promising results and might help getting relative efflux level measurements in Gram- bacteria. However, precise concentrations & timing to use are yet to be determined for each particular bacterial species and efflux pump involved.

To sum up, the applicability of ColorFlux in Gram- bacteria will depend on the intended application. While ColorFlux can be useful as a tool to quickly check for the presence/absence of efflux pumps (i.e. to validate efflux pump knock-out mutants), some protocol optimization will be required if you are aiming at comparing relative efflux activity levels between strains, or to detect resistant bacteria.


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

A recent study utilized the Total Complement Functional Screen ELISA from Svar Life Sciences! Check out the abstract and full text below.


Abstract

Reservoir host associations have been observed among and within Borrelia genospecies, and host complement-mediated killing is a major determinant in these interactions. In North America, only a subset of Borrelia burgdorferi lineages cause the majority of disseminated infections in humans. We hypothesize that differential resistance to human complement-mediated killing may be a major phenotypic determinant of whether a lineage can establish systemic infection. As a corollary, we hypothesize that borreliacidal action may differ among human subjects. To test these hypotheses, we isolated primary B. burgdorferi clones from field-collected ticks and determined whether the killing effects of human serum differed among those clones in vitro and/or whether these effects were consistent among human sera. Clones associated with human invasiveness did not show higher survival in human serum compared to noninvasive clones. These results indicate that differential complement-mediated killing of B. burgdorferi lineages is not a determinant of invasiveness in humans. Only one significant difference in the survivorship of individual clones incubated in different human sera was detected, suggesting that complement-mediated killing of B. burgdorferi is usually similar among humans. Mechanisms other than differential human complement-mediated killing of B. burgdorferi lineages likely explain why only certain lineages cause the majority of disseminated human infections.

Pearson, P.; Rich, C.; Siegel, E.L.; Brisson, D.; Rich, S.M. Differential Resistance of Borrelia burgdorferi Clones to Human Serum-Mediated Killing Does Not Correspond to Their Predicted Invasiveness. Pathogens 2023, 12, 1238. https://doi.org/10.3390/pathogens12101238


If you have any questions about the Total Complement Functional Screen ELISA or any of our other offerings, contact us here.

Eagle Bioscience’s GLP-2 ELISA Assay Kit was highlighted in recent publication! The scientists in this study induced the physiological secretion of GLP-2 via nanoparticles to help them develop a combination therapy for inflammatory bowel disease (IBD). Check out the abstract and full article below.


Abstract

Current treatments for inflammatory bowel disease (IBD) treatment consist of anti-inflammatory products. In this study, we sought to induce the physiological secretion of glucagon-like peptide 2, a peptide with intestinal growth-promoting activity, via nanoparticles while simultaneously providing with immunomodulation by tailoring the nanoparticle surface. To this end, we developed hybrid lipid hyaluronate-KPV conjugated nanoparticles loaded with teduglutide for combination therapy in IBD. The nanocarriers induced (or did not induce) immunosuppression depending on the presence (or absence) of a hyaluronan-KPV functionalization. This strategy holds promise as a nanoparticle platform for combined mucosal healing and immunomodulation in IBD treatment.

V. Marotti, et al. A nanoparticle platform for combined mucosal healing and immunomodulation in inflammatory bowel disease treatment, Bioactive Materials, Volume 32, 2024, Pages 206-221, ISSN 2452-199X, https://doi.org/10.1016/j.bioactmat.2023.09.014.


If you have any questions about the GLP-2 ELISA Assay Kit or any of our other offerings contact us here.

The Eagle Bioscience’s Calprotectin ELISA Assay Kit was utilized in a recent study! This study investigated whether microbiome profiles can indidcate stress reactivity in ulcerative colitis. Check out the abstract and full text below!


Abstract

Background: Stress reactivity (SR) is associated with increased risk of flares in ulcerative colitis (UC) patients. Because both preclinical and clinical data support that stress can influence gut microbiome composition and function, we investigated whether microbiome profiles of SR exist in UC.

Methods: Ninety-one UC subjects in clinical and biochemical remission were classified into high and low SR groups by questionnaires. Baseline and longitudinal characterization of the intestinal microbiome was performed by 16S rRNA gene sequencing and fecal and plasma global untargeted metabolomics. Microbe, fecal metabolite, and plasma metabolite abundances were analyzed separately to create random forest classifiers for high SR and biomarker-derived SR scores.

Results: High SR reactivity was characterized by altered abundance of fecal microbes, primarily in the Ruminococcaceae and Lachnospiraceae families; fecal metabolites including reduced levels of monoacylglycerols (endocannabinoid-related) and bile acids; and plasma metabolites including increased 4-ethyl phenyl sulfate, 1-arachidonoylglycerol (endocannabinoid), and sphingomyelin. Classifiers generated from baseline microbe, fecal metabolite, and plasma metabolite abundance distinguished high vs low SR with area under the receiver operating characteristic curve of 0.81, 0.83, and 0.91, respectively. Stress reactivity scores derived from these classifiers were significantly associated with flare risk during 6 to 24 months of follow-up, with odds ratios of 3.8, 4.1, and 4.9. Clinical flare and intestinal inflammation did not alter fecal microbial abundances but attenuated fecal and plasma metabolite differences between high and low SR.

Conclusions: High SR in UC is characterized by microbial signatures that predict clinical flare risk, suggesting that the microbiome may contribute to stress-induced UC flares.

Jonathan P Jacobs, Jenny S Sauk, Aaron I Ahdoot, Fengting Liang, William Katzka, Hyo Jin Ryu, Ariela Khandadash, Venu Lagishetty, Jennifer S Labus, Bruce D Naliboff, Emeran A Mayer, Microbial and Metabolite Signatures of Stress Reactivity in Ulcerative Colitis Patients in Clinical Remission Predict Clinical Flare Risk, Inflammatory Bowel Diseases, 2023;, izad185, https://doi.org/10.1093/ibd/izad185


If you have any questions about the Calprotectin ELISA or any of our other offerings, contact us here.

The Eagle Bioscience’s Glutathione Total Assay Kit and Malondialdehyde HPLC Assay Kit were highlighted in a recent study! The study was conduct to examine the shielding influence of cinnamic acid against lung fibrosis induced by methotrexate. Check out the abstract and full article below!


Abstract

Purpose: Lung fibrosis is a heterogeneous lung condition characterized by excessive accumulation of scarred tissue, leading to lung architecture destruction and restricted ventilation. The current work was conducted to examine the probable shielding influence of cinnamic acid against lung fibrosis induced by methotrexate.

Methods: Rats were pre-treated with oral administration of cinnamic acid (50 mg/kg/day) for 14 days, whereas methotrexate (14 mg/kg) was orally given on the 5th and 12th days of the experiment. Pirfenidone (50 mg/kg/day) was used as a standard drug. At the end of the experiment, oxidative parameters (malondialdehyde, myeloperoxidase, nitric oxide, and total glutathione) and inflammatory mediators (tumor necrosis factor-α and interleukin-8), as well as transforming growth factor-β and collagen content, as fibrosis indicators, were measured in lung tissue.

Results: Our results revealed that cinnamic acid, as pirfenidone, effectively prevented the methotrexate-induced overt histopathological damage. This was associated with parallel improvements in oxidative, inflammatory, and fibrotic parameters measured. The outcomes of cinnamic acid administration were more or less the same as those of pirfenidone. In conclusion, pre-treatment with cinnamic acid protects against methotrexate-induced fibrosis, making it a promising prophylactic adjuvant therapy to methotrexate and protecting against its possible induction of lung fibrosis.

Abdalhameid, E., Abd El-Haleim, E.A., Abdelsalam, R.M. et al. Cinnamic acid mitigates methotrexate-induced lung fibrosis in rats: comparative study with pirfenidone. Naunyn-Schmiedeberg’s Arch Pharmacol (2023). https://doi.org/10.1007/s00210-023-02652-w


If you have any question about the Glutathione Assay and Malondialdehyde HPLC Kit or any of our other offerings contact us here.

The Eagle Bioscience’s Secretory IgA ELISA Assay Kit was highlighted in a recent publication! This study conducted a double-blind randomized trial to help determine if synbiotic supplementation is effective in healthy adults. Check out the abstract and full text below.


Abstract

Synbiotics are increasingly used by the general population to boost immunity. However, there is limited evidence concerning the immunomodulatory effects of synbiotics in healthy individuals. Therefore, we conducted a double-blind, randomized, placebo-controlled study in 106 healthy adults. Participants were randomly assigned to receive either synbiotics (containing Bifidobacterium lactis HN019 1.5 × 108 CFU/d, Lactobacillus rhamnosus HN001 7.5 × 107 CFU/d, and fructooligosaccharide 500 mg/d) or placebo for 8 weeks. Immune parameters and gut microbiota composition were measured at baseline, mid, and end of the study. Compared to the placebo group, participants receiving synbiotic supplementation exhibited greater reductions in plasma C-reactive protein (P = 0.088) and interferon-gamma (P = 0.008), along with larger increases in plasma interleukin (IL)-10 (P = 0.008) and stool secretory IgA (sIgA) (P = 0.014). Additionally, synbiotic supplementation led to an enrichment of beneficial bacteria (Clostridium_sensu_stricto_1, Lactobacillus, Bifidobacterium, and Collinsella) and several functional pathways related to amino acids and short-chain fatty acids biosynthesis, whereas reduced potential pro-inflammatory Parabacteroides compared to baseline. Importantly, alternations in anti-inflammatory markers (IL-10 and sIgA) were significantly correlated with microbial variations triggered by synbiotic supplementation. Stratification of participants into two enterotypes based on pre-treatment Prevotella-to-Bacteroides (P/B) ratio revealed a more favorable effect of synbiotic supplements in individuals with a higher P/B ratio. In conclusion, this study suggested the beneficial effects of synbiotic supplementation on immune parameters, which were correlated with synbiotics-induced microbial changes and modified by microbial enterotypes. These findings provided direct evidence supporting the personalized supplementation of synbiotics for immunomodulation.

Xiaoqin Li, Shan Hu, Jiawei Yin, Xiaobo Peng, Lei King, Linyan Li, Zihui Xu, Li Zhou, Zhao Peng, Xiaolei Ze, Xuguang Zhang, Qiangchuan Hou, Zhilei Shan & Liegang Liu (2023) Effect of synbiotic supplementation on immune parameters and gut microbiota in healthy adults: a double-blind randomized controlled trial, Gut Microbes, 15:2, DOI: 10.1080/19490976.2023.2247025


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