SepMate™-50 (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: 446 USD

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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™-50 (IVD), 100 Tubes (Catalog #85450)
    • Dispenser box containing 4 bags, 25 Tubes/Bag
  • SepMate™-50 (IVD), 500 Tubes (Catalog #85460)
    • Dispenser box containing 4 bags, 25 Tubes/Bag (Catalog #85450) 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

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

(28)
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.
JCI insight 2019 may

Self-tolerance curtails the B cell repertoire to microbial epitopes.

A. Watanabe et al.

Abstract

Immunological tolerance removes or inactivates self-reactive B cells, including those that also recognize cross-reactive foreign antigens. Whereas a few microbial pathogens exploit these holes" in the B cell repertoire by mimicking host antigens to evade immune surveillance the extent to which tolerance reduces the B cell repertoire to foreign antigens is unknown. Here we use single-cell cultures to determine the repertoires of human B cell antigen receptors (BCRs) before (transitional B cells) and after (mature B cells) the second B cell tolerance checkpoint in both healthy donors and in patients with systemic lupus erythematosus (SLE) . In healthy donors the majority ({\~{}}70{\%}) of transitional B cells that recognize foreign antigens also bind human self-antigens (foreign+self) and peripheral tolerance halves the frequency of foreign+self-reactive mature B cells. In contrast in SLE patients who are defective in the second tolerance checkpoint frequencies of foreign+self-reactive B cells remain unchanged during maturation of transitional to mature B cells. Patterns of foreign+self-reactivity among mature B cells from healthy donors differ from those of SLE patients. We propose that immune tolerance significantly reduces the scope of the BCR repertoire to microbial pathogens and that cross-reactivity between foreign and self epitopes may be more common than previously appreciated."
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.
Nature medicine 2018 OCT

Translational control of tumor immune escape via the eIF4F-STAT1-PD-L1 axis in melanoma.

M. Cerezo et al.

Abstract

Preventing the immune escape of tumor cells by blocking inhibitory checkpoints, such as the interaction between programmed death ligand-1 (PD-L1) and programmed death-1 (PD-1) receptor, is a powerful anticancer approach. However, many patients do not respond to checkpoint blockade. Tumor PD-L1 expression is a potential efficacy biomarker, but the complex mechanisms underlying its regulation are not completely understood. Here, we show that the eukaryotic translation initiation complex, eIF4F, which binds the 5' cap of mRNAs, regulates the surface expression of interferon-$\gamma$-induced PD-L1 on cancer cells by regulating translation of the mRNA encoding the signal transducer and activator of transcription 1 (STAT1) transcription factor. eIF4F complex formation correlates with response to immunotherapy in human melanoma. Pharmacological inhibition of eIF4A, the RNA helicase component of eIF4F, elicits powerful antitumor immune-mediated effects via PD-L1 downregulation. Thus, eIF4A inhibitors, in development as anticancer drugs, may also act as cancer immunotherapies.
Genes {\&} Immunity 2018 MAY

Mutations in RNA Polymerase III genes and defective DNA sensing in adults with varicella-zoster virus CNS infection

M. E. Carter-Timofte et al.

Abstract

Recently, deficiency in the cytosolic DNA sensor RNA Polymerase III was described in children with severe primary varicella-zoster virus (VZV) infection in the CNS and lungs. In the present study we examined adult patients with VZV CNS infection caused by viral reactivation. By whole exome sequencing we identified mutations in POL III genes in two of eight patients. These mutations were located in the coding regions of the subunits POLR3A and POLR3E. In functional assays, we found impaired expression of antiviral and inflammatory cytokines in response to the POL III agonist Poly(dA:dT) as well as increased viral replication in patient cells compared to controls. Altogether, this study provides significant extension on the current knowledge on susceptibility to VZV infection by demonstrating mutations in POL III genes associated with impaired immunological sensing of AT-rich DNA in adult patients with VZV CNS infection.
THIS PRODUCT IS MANUFACTURED UNDER A cGMP QUALITY MANAGEMENT SYSTEM COMPLIANT TO 21 CFR 820 AND CERTIFIED TO ISO 13485. PRODUCTS ARE FOR PROFESSIONAL IN VITRO DIAGNOSTIC USE.