SepMate™-15 (IVD)

Tube for density gradient centrifugation for in vitro diagnostic (IVD) applications

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Tube for density gradient centrifugation for in vitro diagnostic (IVD) applications
From: 361 USD

New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more

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Overview

SepMate™ is now manufactured under cGMP and registered as an In Vitro Diagnostic (IVD) device in Australia, Canada, Europe and USA only. In China SepMate™ is considered a non-medical device by the China Food and Drug Administration (CFDA), and should therefore be used as general laboratory equipment. The end user is responsible for determining whether the product is suitable for their specific application. Due to these changes, users in Australia, Canada, Europe, USA and China should, from now on, order SepMate™-15 (IVD) (Catalog #85415 and #85420) and SepMate™-50 (IVD) (Catalog #85450 and #85460). Users in all other countries please refer to SepMate™-15 (RUO) (Catalog #86415) and SepMate™-50 (RUO) (Catalog #86450). Changes to SepMate™ have no impact on the product composition, performance or protocol.

SepMate™ is a tube that facilitates the isolation of PBMCs or specific cell types by density gradient centrifugation. SepMate™ tubes contain an insert that provides a barrier between the density gradient medium and blood. SepMate™ eliminates the need for careful layering of blood onto the density gradient medium, and allows for fast and easy harvesting of the isolated mononuclear cells with a simple pour. SepMate™ is also compatible with RosetteSep™ enrichment cocktails, allowing isolation of specific cell types in less than 30 minutes.
Advantages:
• Eliminates the need for carefully layering blood over the density gradient medium (e.g. Lymphoprep™, etc.)
• Reduces total centrifuge time to 10 minutes with the brake on for fresh samples
• Allows fast and easy harvesting of the isolated mononuclear cells by simply pouring off the supernatant
• Can be combined with RosetteSep™ enrichment cocktails to isolate specific cell types in just 30 minutes
Components:
  • SepMate™-15 (IVD), 100 Tubes (Catalog #85415)
    • Dispenser box containing 4 bags, 25 Tubes/Bag
  • SepMate™-15 (IVD), 500 Tubes (Catalog #85420)
    • Dispenser box containing 4 bags, 25 Tubes/Bag (Catalog #85415) x 5
Contains:
Polypropylene tube containing an insert
Subtype:
Centrifugation Tubes
Cell Type:
B Cells; Dendritic Cells; Monocytes; Mononuclear Cells; NK Cells; T Cells; T Cells, CD4+; T Cells, CD8+; T Cells, Other Subsets; T Cells, Regulatory
Species:
Human
Sample Source:
Bone Marrow; Cord Blood; Whole Blood
Selection Method:
Negative
Application:
Cell Isolation; In Vitro Diagnostic
Brand:
SepMate
Area of Interest:
Chimerism; HLA; Immunology

Scientific Resources

Product Documentation

Educational Materials

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Data and Publications

Data

PBMC recovery from fresh whole blood using SepMate™-50 versus standard density gradient centrifugation. Graph also shows PBMC recovery from a 48 hour-old sample using SepMate™. n in each group = 7

Figure 1. Recovery of mononuclear cells (MNCs) from peripheral blood using SepMate™-50 versus standard density gradient centrifiguation.

Recovery of MNCs from fresh and 48-hour post blood draw enriched by density gradient centrifugation with SepMate™ (purple) or without (grey). There was no significant difference in the recovery of MNCS with and without SepMate™.

PBMC recovery from fresh whole blood using SepMate™-50 versus standard density gradient centrifugation. Graph also shows PBMC recovery from a 48 hour-old sample using SepMate™. n in each group = 7

Figure 2. Human CD4+ T Cell Isolation using SepMate™-50 and RosetteSep™ Human CD4+ T Cell Enrichment Cocktail

Publications

(23)
European journal of human genetics : EJHG 2019 may

Biallelic variants in POLR3GL cause endosteal hyperostosis and oligodontia.

P. A. Terhal et al.

Abstract

RNA polymerase III (Pol III) is an essential 17-subunit complex responsible for the transcription of small housekeeping RNAs such as transfer RNAs and 5S ribosomal RNA. Biallelic variants in four genes (POLR3A, POLR3B, and POLR1C and POLR3K) encoding Pol III subunits have previously been found in individuals with (neuro-) developmental disorders. In this report, we describe three individuals with biallelic variants in POLR3GL, a gene encoding a Pol III subunit that has not been associated with disease before. Using whole exome sequencing in a monozygotic twin and an unrelated individual, we detected homozygous and compound heterozygous POLR3GL splice acceptor site variants. RNA sequencing confirmed the loss of full-length POLR3GL RNA transcripts in blood samples of the individuals. The phenotypes of the described individuals are mainly characterized by axial endosteal hyperostosis, oligodontia, short stature, and mild facial dysmorphisms. These features largely fit within the spectrum of phenotypes caused by previously described biallelic variants in POLR3A, POLR3B, POLR1C, and POLR3K. These findings further expand the spectrum of POLR3-related disorders and implicate that POLR3GL should be included in genetic testing if such disorders are suspected.
Nature immunology 2019

The immune cell landscape in kidneys of patients with lupus nephritis.

A. Arazi et al.

Abstract

Lupus nephritis is a potentially fatal autoimmune disease for which the current treatment is ineffective and often toxic. To develop mechanistic hypotheses of disease, we analyzed kidney samples from patients with lupus nephritis and from healthy control subjects using single-cell RNA sequencing. Our analysis revealed 21 subsets of leukocytes active in disease, including multiple populations of myeloid cells, T cells, natural killer cells and B cells that demonstrated both pro-inflammatory responses and inflammation-resolving responses. We found evidence of local activation of B cells correlated with an age-associated B-cell signature and evidence of progressive stages of monocyte differentiation within the kidney. A clear interferon response was observed in most cells. Two chemokine receptors, CXCR4 and CX3CR1, were broadly expressed, implying a potentially central role in cell trafficking. Gene expression of immune cells in urine and kidney was highly correlated, which would suggest that urine might serve as a surrogate for kidney biopsies.
Theranostics 2019

Characterization and Therapeutic Application of Mesenchymal Stem Cells with Neuromesodermal Origin from Human Pluripotent Stem Cells.

H. Wang et al.

Abstract

Rationale: Mesenchymal stem cells (MSC) hold great promise in the treatment of various diseases including autoimmune diseases, inflammatory diseases, etc., due to their pleiotropic properties. However, largely incongruent data were obtained from different MSC-based clinical trials, which may be partially due to functional heterogeneity among MSC. Here, we attempt to derive homogeneous mesenchymal stem cells with neuromesodermal origin from human pluripotent stem cells (hPSC) and evaluate their functional properties. Methods: Growth factors and/or small molecules were used for the differentiation of human pluripotent stem cells (hPSC) into neuromesodermal progenitors (NMP), which were then cultured in animal component-free and serum-free induction medium for the derivation and long-term expansion of MSC. The resulted NMP-MSC were detailed characterized by analyzing their surface marker expression, proliferation, migration, multipotency, immunomodulatory activity and global gene expression profile. Moreover, the in vivo therapeutic potential of NMP-MSC was detected in a mouse model of contact hypersensitivity (CHS). Results: We demonstrate that NMP-MSC express posterior HOX genes and exhibit characteristics similar to those of bone marrow MSC (BMSC), and NMP-MSC derived from different hPSC lines show high level of similarity in global gene expression profiles. More importantly, NMP-MSC display much stronger immunomodulatory activity than BMSC in vitro and in vivo, as revealed by decreased inflammatory cell infiltration and diminished production of pro-inflammatory cytokines in inflamed tissue of CHS models. Conclusion: Our results identify NMP as a new source of MSC and suggest that functional and homogeneous NMP-MSC could serve as a candidate for MSC-based therapies.
Molecular nutrition {\&} food research 2019

Human Milk Oligosaccharides Protect against Necrotizing Enterocolitis by Inhibiting Intestinal Damage via Increasing the Proliferation of Crypt Cells.

C. Wang et al.

Abstract

SCOPE Necrotizing enterocolitis (NEC) is a devastating disease that is highly lethal in premature infants. Human milk oligosaccharides (HMOs) efficiently reduce the incidence of NEC. However, the protective mechanism of HMO treatment is unknown. It is hypothesized that HMOs protect against NEC by inhibiting the damage to intestinal epithelial cells. METHODS AND RESULTS C57BL/6 pups are challenged with hypoxia and cold stress to induce NEC. All pups are sacrificed after 72 h. It is found that HMO administration reduces the concentrations of IL-8 in the serum and ileum of all NEC mice. Ileum toll-like receptor 4 (TLR4) protein expression and nuclear factor kappa-B (NF$\kappa$B) pathway activation are inhibited. The proliferative ability of enterocytes in the ileum is restored as determined by labeling with proliferation markers (Ki67, SOX9). In a 3D culture intestinal crypt organoids study, HMO treatment improves the maturation of organoid cells and increases the ratio of proliferative cells under lipopolysaccharides (LPS) treatment. HMO treatment downregulates TLR4 expression in the organoid cells, thus reducing the effect of LPS. CONCLUSION HMOs protect intestinal epithelial cells from injury by accelerating the turnover of crypt cells by reducing the expression of TLR4 on intestinal epithelial cells.
Scientific Reports 2018 SEP

IL-27 amplifies cytokine responses to Gram-negative bacterial products and Salmonella typhimurium infection.

C. Petes et al.

Abstract

Cytokine responses from monocytes and macrophages exposed to bacteria are of particular importance in innate immunity. Focusing on the impact of the immunoregulatory cytokine interleukin (IL)-27 on control of innate immune system responses, we examined human immune responses to bacterial products and bacterial infection by E. coli and S. typhimurium. Since the effect of IL-27 treatment in human myeloid cells infected with bacteria is understudied, we treated human monocytes and macrophages with IL-27 and either LPS, flagellin, or bacteria, to investigate the effect on inflammatory signaling and cytokine responses. We determined that simultaneous stimulation with IL-27 and LPS derived from E. coli or S. typhimurium resulted in enhanced IL-12p40, TNF-$\alpha$, and IL-6 expression compared to that by LPS alone. To elucidate if IL-27 manipulated the cellular response to infection with bacteria, we infected IL-27 treated human macrophages with S. typhimurium. While IL-27 did not affect susceptibility to S. typhimurium infection or S. typhimurium-induced cell death, IL-27 significantly enhanced proinflammatory cytokine production in infected cells. Taken together, we highlight a role for IL-27 in modulating innate immune responses to bacterial infection.
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