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.


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

FluoBolt- KLOTHO Fluorescence Immunoassay

High Sensitivity; Single Step Immunoassay for α-KLOTHO in Human Serum and Plasma

Eagle Biosciences is excited to highlight the FloBolt- KLOTHO Fluorescence Immunoassay from Fianostics! This assay detects only α-KLOTHO and does not cross-react with β-KLOTHO. No interference of recombinant FGF-23 with the assay’s signal up to a 100-fold molar excess was monitored. Check out the assay background and highlights below!


Background

α-KLOTHO is expressed in kidney, small intestine, placenta and prostate. The soluble peptide can be found in serum and cerebrospinal fluid. It may play a role in the calcium/ phosphorus homeostasis regulation by e.g. inhibiting active vitamin D synthesis. Further, it is also known as an antiaging-hormone by extending life span by inhibiting insulin/ IGF1 signaling pathway, as experiments in mice showed. KLOTHO is a co-receptor of fibroblast growth factor 23 (FGF-23). Research has investigated association of altered serum KLOTHO levels with chronic kidney disease and failure, renal and hepatocellular carcinomas, osteoporosis or cardiovascular diseases.

α-KLOTHO can be found either as a membrane bound or a secreted form. The membrane bound form consists of 1012 amino acids (aa), starting with a 56 aa long signaling sequence and followed by two glycosyl hydrolase 1 regions (position 57-506 and 515-953). Both glycosyl hydrolase 1 regions lack one essential Glu active site residue. Thus, it is inactive in vivo as a glycosidase although it belongs to the glycosyl hydrolase 1 family. KLOTHO’s secreted isoform, which predominates over the membrane bound form, consists of 549 amino acids (aa). It is produced by alternative splicing and differs from the membrane bound form by aa 550 to 1012 missing.


FluoBolt- KLOTHO Fluorescence Immunoassay Highlights

  • High Sensitivity
  • Single Step Procedure
  • No Washing Steps
  • No Enzyme Substrate required
  • Long Term Stable Signal

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


We are excited to bring you the new Rat NT-proBNP ELISA Assay Kit! This product has been developed, validated, and manufactured by Biomedica in Austria. This assay extends our robust line of kits specific to cardiac biomarkers. Learn more below!


Biomarker Background

BNP is mainly expressed by the ventricular myocardium in response to volume overload and increased filling pressure. It is synthesized and secreted by cardiomyocytes. Mature BNP consists of 108 amino acids (proBNP or BNP-108). ProBNP is cleaved during secretion in a 1:1 ratio resulting in physiologically active BNP-32 and the biologically inactive 76 amino acid NT-proBNP. NT-proBNP (1-76) has greater plasma stability and a much longer biological half-life (90-120 minutes) than BNP, being considered as the preferred laboratory marker. BNP has a key role in cardiovascular homeostasis with biological actions including natriuresis, diuresis, vasorelaxation, and inhibition of renin and aldosterone secretion. A high concentration of BNP in the bloodstream is indicative of heart failure.


Rat NT-proBNP ELISA Assay Kit Features

  • Low Sample Volume Required – 10 µL/well
  • Controls Included
  • Sample Values Provided

Related Products

NT-proBNP ELISA Assay Kit
NT-proCNP ELISA Assay Kit
NT-proANP ELISA Assay Kit
BNP Fragment ELISA Kit


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

Fetuin A (PTM) ELISA New Product Announcement

Eagle Biosciences is excited to partner with BPM and support the Fetuin A (PTM) ELISA (DNlite-DKD)!

About Fetuin A (PTM)

The unique Fetuin A post translation modifications measured in this assay were identified in a large-scale profiling of urinary proteomics. This new biomarker can help predict the kidney condition of diabetes patients, months to years in advanced. This urine test can help predict kidney decline or complications and potentially improve a patient with diabetic kidney disease quality of care.

Principle of the Fetuin A (PTM) ELISA

The Fetuin A (PTM) ELISA (unique Fetuin-A with specific post translational modification (PTM) for Diabetic Kidney Disease (DKD)) is a competitive immunoassay. In this Fetuin A (PTM) ELISA, calibrators or unknown urine samples are mixed with anti-unique PTM Fetuin-A monoclonal antibody (mAb), and then incubated in a microplate pre-bounded with unique PTM Fetuin-A. The monoclonal antibody recognizes unique PTM Fetuin-A in calibrators or unknown samples under competition in microplate wells. After an incubation, an Horseradish Peroxide (HRP) conjugated secondary antibody is added, followed by an incubation with 3,3’,5,5’-tetramethylbenzidine (TMB) substrate. Their relative reactivity is determined by absorbance measurement at 450 nanometers (nm) and plotted by comparison with a predetermined unique PTM Fetuin-A calibration curve.

Benefits of the Assay

    • Fewer Steps
    • Shorter processing times – ever for high-throughput samples

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

Cortisol ELISA Assay Kit

Eagle Biosciences is excited to announce the new Cortisol Saliva ELISA Assay Kit. The kit is intended for the quantitative determination of cortisol in human saliva.


Why Measure Cortisol?

Cortisol is the most abundant circulating steroid and the major glucocorticoid secreted by the adrenal cortex. Cortisol is physiologically effective in blood pressure maintenance and anti-inflammatory activity. It is also involved in calcium absorption, gluconeogenesis as well as the secretion of gastric acid and pepsin. It is increased under stress situations, physical exercise and external administration of ACTH. Most circulating cortisol is bound to cortisol binding globulin or transcortin and albumin. The free cortisol, which is considered the active part of blood, is about 1–2%. In the absence of appreciable amounts of the cortisol binding proteins in saliva, salivary cortisol is considered to be free and shows a diurnal rhythm with the highest levels in the morning and the lowest levels at night.

Measurement of cortisol levels in general can be used as an indicator of adrenal function and the differential diagnosis of Addison’s and Cushing’s diseases as well as adrenal hyperplasia and carcinoma.


Check out the benefits of Cortisol Saliva ELISA Assay Kit:

  • NO SHAKING required
  • ROOM TEMPERATURE incubation
  • SMALL SPECIMEN SAMPLE required

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

Anti-tTG sIgA/IgA ELISA Assay Kit

Eagle Biosciences is excited to announce the new Anti-tTG sIgA/IgA ELISA Assay Kit. This kit is intended for the quantitative determination of anti-human tissue transglutaminase sIgA/IgA in stool samples.

Assay Background


Celiac disease is a chronic illness of the small intestinal mucous membrane. The reason is an intolerance against gluten, which is found in many cereals. This food intolerance against cereal proteins leads to a production of auto-antibodies against tissue transglutaminase, which is the major antigen for endomysium antibodies.

The intake of gluten-containing food leads to an inflammation of the small intestinal mucous membrane, which results in a reduced absorption of nutrients. The symptoms of the disease are reduction of weight, diarrhea, vomitus, anorexia and tiredness. The growth of children is impaired. The characteristic of the symptoms might be different. The only therapeutic treatment is a gluten-free diet. A non-treated celiac disease increases the risk of non-Hodgkin-lymphoma and colon cancer. Celiac disease is associated with type 1 diabetes mellitus in five to ten percent of the patients. Women are more often affected than men. The outcome of the disease is pronounced during infancy and in ages between 30 and 40 years old.

Applications:

  • Food Intolerance
  • Monitoring an Elimination Diet

Anti-tTG sIgA/IgA ELISA Assay Kit Advantages

  • Easy
  • Rapid
  • Precise
  • All reagents included

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

New Insulin ELISA Assay Kits

Eagle Bioscience’s has extended their line of insulin assays! We now offer a wide range of Insulin ELISA Assay Kits that measure insulin concentration in a number of different species samples. These new kits include the Ovine Insulin ELISA Assay Kit, Equine Insulin ELISA Assay Kit, Porcine Insulin ELISA Assay Kit, Canine Insulin ELISA Assay Kit. These kits are highly sensitive, reliable, and easy to use.

More about insulin:

Insulin is a peptide hormone, produced by beta cells of the pancreas. Enzymatic cleavage of proinsulin results in the production of equimolar amounts of insulin and C-peptide in circulation. Insulin is central to regulating carbohydrate and fat metabolism in the body.  Excessive amounts of insulin are associated with excess amounts of glucose in the system. High levels of insulin are known to cause weight gain, water bloating, high blood pressure, magnesium deficiency and an increase in the amount of inflammatory compounds in the blood, which causes blood clots and blood vessel damage. Insulin, when insufficiently produced, results in diabetes mellitus. In most cases, a high fasting insulin level is consistent with insulin resistance symptoms, but in some cases, it can be caused by more serious conditions such as Cushing’s syndrome, acromegaly or possibly insulinoma.

Related Insulin ELISA Assay Kits:

Proinsulin ELISA Assay Kit

Insulin ELISA Assay Kit

Insulin Ultrasensitive ELISA Assay Kit

If you are looking for any other specific related products, or have any questions about our offerings contact us here.

Introducing the FGF-19 ELISA Assay Kit and the Mouse FGF-21 ELISA Kit. These kits are an extension of an expansive FGF Assay line of kits Eagle Biosciences already offers. Fibroblast Growth Factors are a family of cell signaling proteins that are involved in a wide variety of biological processes. FGF-19 and FGF-21 belong to a subfamily of these proteins that function as an important role in nutrient metabolism.

More about FGF-19:

The primary source of endocrine FGF-19 is the ileum, bile acids release into the intestine after a meal to induce expression of FGF-19. Circulating FGF-19 plays an important role in maintaining proper bile acid homeostasis. Several pharmacologic studies in hyperglycemic, obese animal models have shown that FGF-19 can improve metabolic rate and lower serum glucose and hepatic triglyceride and cholesterol levels. Like insulin, FGF-19 functions as postprandial hormone to govern hepatic protein synthesis, glycogen synthesis and gluconeogenesis, but does not stimulate lipogenesis.

More about FGF-21:

FGF-21 has been implicated in the regulation of lipid and glucose metabolism under fasting and ketotic conditions. In murine models, FGF-21 is predominantly expressed in liver, but it also expressed in adipose tissue and pancreatic β-cells. FGF-21 stimulates glucose uptake in adipocytes. It also protects animals from diet-induced obesity when overexpressed in transgenic mice and lowers blood glucose and triglyceride levels when administered to diabetic rodents. When administered daily for 6 weeks to diabetic rhesus monkeys, FGF-21 caused a dramatic decline in fasting plasma glucose, fructosamine, triglycerides, insulin, and glucagon. Furthermore, elevated plasma FGF-21 concentrations in humans appear to be related to the presence of hepatic and peripheral insulin resistance.

Related Products:

FGF-23 C-Terminal ELISA Kit

MedFrontier Intact FGF-23 Assay

FGF Basic ELISA Assay

Intact FGF-21 ELISA Kit

Total FGF-21 ELISA Assay

cTerminal FGF-21 ELISA Kit

N-Terminal FGF-21 ELISA Kit

iLite FGF-21 Assay Ready Cells

If you are looking for any other specific related products or if you have questions about our offerings, contacts us here.

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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LEUCINE-RICH ALPHA-2-GLYCOPROTEIN (LRG) ELISA Now Available!

Austrian supplier Biomedica has released a new ELISA, the Leucine-Rich Alpha-2-Glycoprotein or LRG for short. The LRG ELISA Assay kit is intended for the quantitative determination of human LRG in serum, EDTA plasma, heparin plasma, and citrate plasma.

Human LRG ELISA highlights:

Size: 1×96 wells
Sensitivity: 0.26 ng/ml
Standard Range: 0 – 64 ng/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma
Sample Size: 100 µl pre-diluted sample / well (5 µl sample)

About LRG

LRG (leucine-rich alpha-2-glycoprotein) is a glycoprotein with a molecular mass of 38.2 kDa. It is encoded by the human gene LRG-1 which is mapped on chromosome 19 at the cytogenetic band 19p13.3. The protein LRG (or also named LRG1) runs at approximately 50 kDa under reducing conditions, as it contains a carbohydrate content of 23%. LRG is the founding member of the family of leucine-rich repeat proteins. The mature protein consists of 312 amino acids, from Val36 to Gln347, with a leucine content of 66 amino acids. LRG is folded to eight leucine-rich repeat (LRR) domains of 22 amino acid length, and a C-terminal LRRCT domain with 49 amino acid length. Human LRG shows 62.5% sequence identity with mouse LRG, and 60.7% with rat LRG.

LRG binds to the TGFβ accessory receptor endoglin, and in the presence of TGFβ1 this leads to the induction of the TβRII-ALK1-Smad1/5/8 signaling pathway. TGFβ1 therefore promotes binding of LRG to the proangiogenic ALK1 but inhibits the interaction with angiostatic ALK5. Induced signaling leads to endothelial cell proliferation and blood vessel outgrowth.

Like many other family members of the leucine-rich repeat (LRR) family, LRG has multiple binding partners. LRG directly interacts with the mitochondrial electron transfer protein cytochrome c, whereas the physiological relevance of this interaction is not yet known. LRG further binds to TGFβ1, the most frequently expressed TGFβ isoform.

The tissue distribution of LRG varies, with high-level expression in the liver, lower expression in the heart, and minimal expression in spleen and lung. LRG is expressed during hematopoiesis. It plays a role in the innate immune system as it is upregulated during neutrophil differentiation; LRG is packed into peroxidase-negative granules of human neutrophils and then secreted upon activation to modulate the microenvironment. Differential expression of LRG is further associated with certain carcinomas, neurodegenerative disease, aging or autoimmune disease. In addition, studies have demonstrated an association between cardiac remodeling (hypertrophy, fibrosis, abnormal vasculature, heart failure) and reduced expression of LRG.

LRG is involved in cell proliferation and immune response, in cell migration, neovascularization and apoptosis. It is a proangiogenic factor which is involved in the regulation of the TGFβ signaling pathway. Up-regulation of LRG is described in response to acute phase response in hepatocytes.

LRG is potentially a biomarker for a variety of diseases e.g. as inflammatory biomarker for autoimmune diseases such as rheumatoid arthritis and inflammatory bowel disease. Numerous groups have shown that LRG is increased in various immune-related diseases such as psoriasis, juvenile idiopathic arthritis, Kawasaki disease, appendicitis, and cancers, indicating that LRG elevation is not only limited to autoimmune diseases. In addition, LRG may serve as a biomarker for several other disease conditions such as heart failure, and diabetes-related complications. Plasma Leucine-Rich α-2-Glycoprotein has also been demonstrated to predict cardiovascular disease risk in end-stage renal disease. Leucine-rich α-2-glycoprotein is highly expressed in the brain and it is possible to distinguish idiopathic normal pressure hydrocephalus (iNPH) from other neurodegenerative diseases such as Alzheimer disease by measuring LRG in cerebrospinal fluid.

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