Mouse Rat 25-OH Vitamin D ELISA Highlighted in Recent Publication

The Eagle Bioscience’s Mouse Rat 25-OH Vitamin D ELISA was recently highlighted about how the MicroRNA-122 contributes to lipopolysaccharide-induced acute kidney injury via down-regulating the vitamin D receptor in the kidney.

Abstract


Background
Our previous studies showed that vitamin D receptor (VDR) depletion promotes lipopolysaccharide (LPS)-induced acute kidney injury (AKI) in mice, and renal VDR is down-regulated in AKI, but the mechanism of VDR down-regulation is unclear.

Methods
Nutritional vitamin D deficiency was induced by feeding mice a vitamin D-deficient (VD-D) diet. Mice were injected intraperitoneally with LPS (20 mg/kg) to establish LPS-induced AKI. Levels of VDR and miR-122 were measured both in vivo and in vitro. The associations between VDR and miR-122 were analysed by dual-luciferase reporter assays.

Results
Compared with vitamin D-sufficient (VD-S) mice, VD-D mice developed more severe renal injury following LPS challenge. LPS induced a dramatic decrease in VDR expression and marked induction of miR-122 both in vivo and in vitro. Furthermore, miR-122 hairpin inhibitor alleviated LPS-induced VDR down-regulation whereas miR-122 mimic directly suppressed VDR expression in HK-2 cells. In luciferase reporter assays, miR-122 mimic was able to suppress luciferase activity in 293T cells co-transfected with a luciferase reporter that contains a putative miR-122 target site from 3′UTR of the VDR transcript, but not when this site was mutated. Moreover, miR-122 mimic significantly blocked paricalcitol-induced luciferase activity in 293T cells co-transfected with a VDRE-driven luciferase reporter, whereas miR-122 hairpin inhibitor enhanced paricalcitol’s activity to suppress PUMA and caspase 3 activation induced by LPS in HK-2 cells.

Conclusions
Collectively, these studies provide evidence that miR-122 directly targets VDR in renal tubular cells, which strongly suggest that miR-122 up-regulation in the kidney under LPS challenge contributes to kidney injury by down-regulating VDR expression.

He, J., Du, J., Yi, B., et al. MicroRNA-122 contributes to lipopolysaccharide-induced acute kidney injury via down-regulating the vitamin D receptor in the kidney. European Journal of Clinical Investigation. (2021) Full Text Here.


If you have any questions about the Mouse Rat 25-OH Vitamin D ELISA or our other offerings, contact us here.

COVID-19 IgM

The Eagle Bioscience’s Coronavirus COVID-19 IgM ELISA Assay Kit was recently highlighted in a publication about a review on current diagnostic techniques for COVID-19. This review includes techniques such as RT-PCR, ddPCR, LAMP, CRISPR, and immunoassay techniques. To learn more about this publication, read below:

Abstract


Introduction
SARS-Cov-2 first appeared in Wuhan, China, in December 2019 and spread all over the world soon after that. Given the infectious nature of SARS-CoV-2, fast and accurate diagnosis tools are important to detect the virus. In this review, we discuss the different diagnostic tests that are currently being implemented in laboratories and provide a description of various COVID-19 kits.

Areas Covered
We summarize molecular techniques that target the viral load, serological methods used for SARS-CoV-2 specific antibodies detection as well as newly developed faster assays for the detection of SARS-CoV-2 in various biological samples.

Expert Opinion
In the light of the widespread pandemic, the massive diagnosis of COVID-19, using various detection techniques, appears to be the most effective strategy for monitoring and containing its propagation.

Jaddaoui, IE., Allali, M., Raoui, S., et al. A review on current diagnostic techniques for COVID-19. Expert Review of Molecular Diagnostics. (2021) 21:2, 141-160.


If you have any questions about this Coronavirus COVID-19 IgM ELISA Assay Kit or our other products, contact use here!

The Eagle Bioscience’s TNF-Alpha ELISA Assay Kit was recently utilized in a publication about the detection rate and genotyping of Cryptosporidium spp. and its relation to corporate TNF-Alpha in elderly Egyptians.

Abstract


Background
Elderly individuals are considered an at-risk population, susceptible to enteric infections; and Cryptosporidium spp. is an apicomplexan protozoan considered to be one of the most common protozoa causing diarrhea. Cryptosporidiosis causes elevation of many pro-inflammatory cytokines like tumor necrosis factor-alpha (TNF-α) which may play a role in pathogenesis of the disease.

Objectives
This study was designed for detection and genotyping of Cryptosporidium spp. in elderly patients and the relationship of infection with copro TNF-α. Diagnosis was by evaluation of permanent acid-fast cold Kinyoun’s (AF) staining, immunochromatographic detection (ICT), and ELISA in comparison to molecular diagnosis as gold standard diagnostic method.

Subjects and Methods
Stool samples were collected from 270 elderly patients aged above 60 years old attending outpatient clinics of Internal Medicine Hospital, Cairo University. Sample were examined microscopically by direct wet mount, and AF straining, and then subjected to ICT ELISA and nested PCR (nPCR) assays. Positive samples by nPCR were then subjected to Restriction fragment length polymorphism (RELP) to detect Cryptosporidium genotypes. Corpo-levels of TNF-Alpha were measured to asses their relationship with cryptosporidiosis.

Results
Cryptosporidiosis detection rates of 3.7%, 6.3%, 6.7%, 3.7% were determined by microscopic examination after AF staining, ICT, ELISA and nPCR, respectively. When RFLP was performed on nPCR positive samples, eight and two samples were assigned as genotype 1 and 2, respectively. Moreover, TNF-α was significantly correlated with cryptosporidiosis.

Conclusion
Conclusion: The elderly are highly vulnerable to cryptosporidiosis. Immunodiagnosis and molecular techniques are fundamental for the diagnosis of cryptosporidiosis. Cryptosporidiosis significantly affects copro TNF-α.

Amin, NM., Raafat, A., Morsy, SM. Detection rate and genotyping of Cryptosporidium spp. and its relation to copro TNF-α in elderly Egyptians attending outpatient clinics of Cairo University Hospitals. Parasitologists. (2021)14:77-85


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Anti-Infliximab ELISA

The Eagle Bioscience’s Anti-Infliximab ELISA Assay Kit was recently highlighted in a publication about factors that are associated with reduced infliximab exposure in the treatment of pediatric autoimmune disorders.

Abstract


Background
Inadequate systemic exposure to infliximab (IFX) is associated with treatment failure. This work evaluated factors associated with reduced IFX exposure in children with autoimmune disorders requiring IFX therapy.

Methods
In this single-center cross-sectional prospective study IFX trough concentrations and anti-drug antibodies (ADAs) were measured in serum from children diagnosed with inflammatory bowel disease (IBD) (n = 73), juvenile idiopathic arthritis (JIA) (n = 16), or uveitis (n = 8) receiving maintenance IFX infusions at an outpatient infusion clinic in a tertiary academic pediatric hospital. IFX concentrations in combination with population pharmacokinetic modeling were used to estimate IFX clearance. Patient demographic and clinical data were collected by chart review and evaluated for their relationship with IFX clearance.

Results
IFX trough concentrations ranged from 0 to > 40 μg/mL and were 3-fold lower in children with IBD compared to children with JIA (p = 0.0002) or uveitis (p = 0.001). Children with IBD were found to receive lower IFX doses with longer dosing intervals, resulting in dose intensities (mg/kg/day) that were 2-fold lower compared to children with JIA (p = 0.0002) or uveitis (p = 0.02). Use of population pharmacokinetic analysis to normalize for variation in dosing practices demonstrated that increased IFX clearance was associated with ADA positivity (p = 0.004), male gender (p = 0.02), elevated erythrocyte sedimentation rate (ESR) (p = 0.02), elevated c-reactive protein (CRP) (p = 0.001), reduced serum albumin concentrations (p = 0.0005), and increased disease activity in JIA (p = 0.009) and IBD (p ≤ 0.08). No significant relationship between diagnosis and underlying differences in IFX clearance was observed. Multivariable analysis by covariate population pharmacokinetic modeling confirmed increased IFX clearance to be associated with anti-IFX antibody positivity, increased ESR, and reduced serum albumin concentrations.

Conclusions
Enhanced IFX clearance is associated with immunogenicity and inflammatory burden across autoimmune disorders. Higher systemic IFX exposures observed in children with rheumatologic disorders are driven primarily by provider drug dose and interval selection, rather than differences in IFX pharmacokinetics across diagnoses. Despite maintenance IFX dosing at or above the standard recommended range for IBD (i.e., 5 mg/kg every 8 weeks), the dosing intensity used in the treatment of IBD is notably lower than dosing intensities used to treat JIA and uveitis, and may place some children with IBD at risk for suboptimal maintenance IFX exposures necessary for treatment response.

Funk, RS., Shakhnovich, V., Cho, YK., et al. Factors associated with reduced infliximab exposure in the treatment of pediatric autoimmun disorders: a cross-sectional prospective convenience sampling study. Pediatric Rheumatology. (2021) 19:62


If you have any questions about the Anti-Infliximab ELISA Assay Kit or our other offerings, contact us here.

The Eagle Bioscience’s Calprotectin ELISA Assay Kit was highlighted in a recent study! In this publication scientists studied the dose effect of bovine lactoferrin (bLF) fortification on diarrhea and respiratory tract infections in weaned infants with anemia.

Abstract


Objective
The aim of this study was to explore the dose effect of bovine lactoferrin (bLF) fortification on the morbidity of diarrhea and respiratory tract infections in weaned infants with anemia.

Methods
A total of 108 infants with anemia, who were exclusively breast fed at 4 to 6 months and weaned and formula fed at 6 to 9 months, were recruited. The eligible infants were randomly assigned to fortified group 0 (FG0), fortified group 1 (FG1), or fortified group 2 (FG2) and were given formula fortified with 0 mg/100 g, 38 mg/100 g, and 76 mg/100 g of bLF, respectively, for 3 mo. The morbidity of diarrhea and respiratory tract infections (RTIs), the duration of respiratory and diarrhea-related illnesses, and the levels of fecal human beta-defensin 2 (HBD-2), cathelicidin LL-37 (LL-37), secretory IgA (sIgA), butyrate, and calprotectin were assessed.

Results
After the exclusion of 12 dropouts, the primary outcome measures, including episodes and duration of diarrhea and RTIs during the intervention, were obtained from 96 infants (35, 33, and 28 in FG0, FG1, and FG2, respectively). Compared with infants in FG0, there was a lower morbidity of rhinorrhea, wheezing, and skin rash among infants in FG1 (P < 0.05) and a lower morbidity of respiratory-related illness and wheezing among infants in FG2 (P < 0.05). Furthermore, a lower morbidity of diarrhea-related illness, diarrhea, vomiting, and nausea was observed among infants in FG2 than those in the other two groups (P < 0.05). In addition, the FG1 infants had a lower morbidity of vomiting and nausea than the FG0 infants (P < 0.05). The HBD-2, LL-37, sIgA, and calprotectin levels were significantly higher whereas the butyrate level was significantly lower in the FG2 infants than in infants in the other two groups after 3 mo of intervention (P < 0.05).

Conclusions
The bLF-fortified formula was effective in reducing the morbidity of diarrhea and RTIs in infants with anemia, with the 76 mg/100 g bLF-fortified formula exhibiting a stronger effect. The bLF fortification could be a new strategy for the prevention of diarrhea and RTIs in infants with anemia.

Chen, K., Jin, S., Chen, H.,et al. Dose effect of bovine lactoferrin fortification on diarrhea and respiratory tract infections in weaned infants with anemia: A randomized, controlled trial. Nutrition. (2021)90:111288


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Studies Show that Neuropilin-1 Could Play Role in Transmission of COVID-19

Eagle News

Two Independent Research Teams Find Possible Link Between Neuropilin-1 and SARS-CoV-2 Transmission

Neuropilin-1 (NRP1) is an essential transmembrane cell surface receptor acting primarily as a co-receptor for various ligands (i.e. VEGF, Semaphorins). Due to alternative splicing or shedding, the extracellular region can be released into circulation as soluble Neuropilin-1. NRP1 functions in many key biological processes including the neuronal, cardiovascular and the immune system. The virus SARS-CoV-2 is the causative agent of the coronavirus disease COVID-19 NRP1 is expressed in multiple cell types in the body but occurs primarily on cells in the lung, nose and brain (i.e. respiratory and olfactory epithelium as well as in the CNS). SARS CoV 2 enters the host cells by its spike proteins mainly through its binding to the angiotensin converting enzyme 2 (ACE2).

Click here for summary of findings.

Studies:

Neuropilin-1 facilitates SARS-CoV-2 cell entry and provides a possible pathway into the central nervous system. Cantuti-Castelvetri L et al., Science 13 Nov, 2020; Vol. 370, Issue 6518, pp. 856-860. Full publication.

Neuropilin-1 is a host factor for SARS-CoV-2 infection. Daly JL et al., Science, 13 Nov 2020; Vol. 370, Issue 6518, pp. 861-865. Full publication.

About the Neuropilin-1 ELISA:

  • Only assay that measures free and ligand bound soluble NRP1
  • Low sample volume – only 10µl needed
  • Highly specific using epitope mapped antibodies
  • Rigorously validated according to international guidelines

Contact us for more information about this or our other SARS-CoV-2 related products.

Eagle Biosciences is now offering a new Human VEGF ELISA Kit from Biomedica. The VEGF ELISA Assay Kit is intended for the quantitative determination of human VEGF in serum, EDTA plasma, and citrate plasma.

VEGF ELISA Highlights:

  • DAY Test – results in 4.5 h
  • High sensitivity – measurable values in both serum and plasma
  • RELIABLE- rigorously validated according to FDA/ICH/EMA guidelines
  • READY to use – calibrators and controls included
  • EXCELLENT correlation to existing methods
  • SMALL sample size – only 10µl sample / well required

Areas of interest:

  • Cancer
  • Metabolic disease (diabetes and diabetic kidney disease, diabetic retinopathy, obesity)
  • Retinal Diseases
  • Autoimmune & inflammatory disease (rheumatoid arthritis, psoriasis, psoriatic arthritis)
  • Heart and cardiovascular disease
  • Skeletal bone formation and bone repair

About VEGF

Vascular endothelial growth factor (VEGF or VEGF-A), is a growth hormone secreted by endothelial cells, fi broblasts, smooth muscle cells, platelets, macrophages, and many other cell types. It belongs to the cysteine-knot growth factor superfamily (1) and has a molecular weight of about 40 kDa. Currently, 17 different VEGF isoforms have been described to be expressed from one single gene. They are produced by alternative promoter usage/initiation or alternative splicing/proteolysis after protein translation. The N-terminal region is responsible for receptor binding and conserved among all VEGF isoforms. In contrast, residues of the C-terminus differ between isoforms and determine protein length and properties: binding to co-receptor Neuropilin-1 (NRP1) or to extracellular matrix (ECM), agonist/antagonist of angiogenesis. Most isoforms result from the common transcripts: VEGF111, VEGF121, VEGF145, VEGF165, VEGF189 and VEGF206. Additionally, a third VEGF variant (VEGFAx), that demonstrates pro- and anti-apoptotic properties, was described. Thus, vascularization is tightly controlled by the balance of various splice variants, their availability and concentration, whereas isoforms linked to the ECM constitute a reservoir of VEGF that can quickly be shed to circulating forms. One of the most potent pro-angiogenic isoforms is VEGF165a. After secretion, 50-70% of VEGF165a is attached to the extracellular matrix (via heparin binding site), the rest is freely diffusible. It is the most abundant isoform and enhances signaling over the VEGFR2 receptor by additionally binding to its co-receptor Neuropilin-1. VEGF A isoforms are glycosylated, homodimeric proteins. Two anti-parallel monomers are linked by intermolecular disulfide bonds whereas eight cysteine residues form a knot-like structure at one end of each monomer. However, heterodimerization with PLGF has been described as well.

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Eagle Biosciences Diagomix

Eagle Biosciences, Inc. is pleased to partner with New York based Diagomix to offer a new line of monoclonal antibodies.

About Diagomix

Diagomix develops and manufactures antibodies, products for immunohistochemistry, ELISA, lateral flow assays and other antibody-based tests. Founded by a team of biomedical professionals, they offer a wide range of biochemical product development services. Their areas of focus include immunohistochemical diagnostics, companion diagnostics, food safety, and hapten detection.

New Product Offerings

Mouse Monoclonal Anti-CHID1 Clone 3D4
Mouse Monoclonal Anti-CD31 Antibody Clone PBM-6B3
Mouse Monoclonal Anti-PCNA Antibody Clone Р60
Mouse Monoclonal Anti-PCNA Antibody Clone Р56
Mouse Monoclonal Anti-Thyroglobulin Antibody Clone 2H11-D5
Mouse Monoclonal Anti-PSA Antibody Clone P1
Mouse Monoclonal Anti-MLH1 Antibody Clone PBM-5A6
Mouse Monoclonal Anti-EpCAM Antibody Clone VU-1D9
Mouse Monoclonal Anti-Desmin Antibody Clone PBM-3B4
Mouse Monoclonal Anti-Pan Cytokeratin Antibody Clone C11
Mouse Monoclonal Anti-Cytokeratin 5 Antibody Clone PBM-2C6
Mouse Monoclonal Anti-CD10 Antibody Clone PBM-1E2
Mouse Monoclonal Anti-Cytokeratin 18 Antibody Clone DC-10
Mouse Monoclonal Anti-AMACR Antibody Clone PBM-G8
Mouse Monoclonal Anti-p53 Antibody Clone BP-53-12
Mouse Monoclonal Anti-ER Antibody Clone PBM-1H7
Mouse Monoclonal Anti-Ki67 Antibody Clone PBM-20G3

Contact us for more information about Diagomix products.

Eagle News

Immune Monitoring COVID-19

Cells infected with SARS-CoV-2 are only visible to CD8+ T cell after virus proteins have been processed and presented by MHC class I molecules – and only then are the CD8+ T cell able to eradicate the infection. While antibodies against the COVID-19 virus are important to prevent or minimize infection, CD8+ T cells are responsible for clearing the virus from the body. Hence, the identification of peptides being presented by MHC class I is essential to:

  • Design potent vaccines eliciting a persistent response and
  • Using MHC class I tetramers to monitor cellular immune responses
  • Assessments of vaccine induced cellular immunity

immunAware allows researchers pursuing novel COVID-19 vaccines to generate tetramers and monitor immune responses. In the current pandemic scenario it is essential not to only focus on a few ALA allotypes; rather it is essential to be able to monitor as many allotypes as possible to ensure a novel vaccine targets a broad range of allotypes.

The easYmer reagents can also be used to validate the binding of predicted epitopes to further stratify the selection of potential COVID-19 vaccine targets.

As the US distributor for immunAware, Eagle Biosciences offer their extensive catalog of MHC tetramer products, including:

HLA-A*29:02 (WYMWLGARY) Class I Tetramer

See our full list of immunAware products here. If you have questions or for assistance with a specific product, please contact us.

Alzheimer’s targets brain cells that help people stay awake

Science News

New Alzheimer’s disease research is providing valuable insight into the cause of the drowsiness associated with dementias. A study performed in a neuropathology lab at UC San Francisco found the brain stem and hypothalamus to be packed with tau protein. The brain stem and hypothalamus oversee several nervous system functions, including keeping people awake and focused. Tau forms “tangles” inside of nerve cells and hinder them from performing necessary functions. In studying three areas of theses structures that help keep people awake, researchers found over 70 percent of the nerve cells were destroyed by tau. These findings will help to drive a greater focus on the brain stem in dementia and Alzheimers research.

Read the full article here.

Eagle Biosciences offers a sensitive and reliable assay for detection of tau in cell culture supernatant, serum, cell culture extracts, plasma, cerebral spinal fluid. We also offer several Tau monomer and preformed fibril products.

Tau ELISA Assay Kit

Active Human Recombinant Tau (K18), P301L mutant Protein Monomer

Active Human Recombinant Tau441 (2N4R) mutant Protein Monomer

Active Human Recombinant Tau (K18), P301L mutant PPF

Active Human Recombinant Tau441 (2N4R), P301S mutant PPF

If you are looking for a specific tau protein related product or if you have questions about our offerings, contact us here.