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RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD56

Immunodensity negative selection cocktail

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From: 403 USD

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Immunodensity negative selection cocktail
From: 403 USD

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Overview

The RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD56 is designed to enrich circulating epithelial tumor cells from fresh whole blood by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD3, CD14, CD16, CD19, CD38, CD45, CD56, CD61, CD66b and glycophorin A on red blood cells (RBCs). When centrifuged over a density gradient medium such as Lymphoprep™ (Catalog #07801), the unwanted cells pellet along with the RBCs. The purified tumor cells are present as a highly enriched population at the interface between the plasma and the density gradient medium.

This cocktail is compatible with breast cancer samples. If you are working with small-cell carcinoma samples, including lung cancer samples, use the RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD36 (Catalog #15127).
Advantages:
• Fast and easy-to-use
• Requires no special equipment or training
• Untouched, viable cells
• Can be combined with SepMate™ for consistent, high-throughput sample processing
Components:
  • RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD56 (Catalog #15137)
    • RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD56, 2 mL
  • RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD56 (Catalog #15177)
    • RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD56, 5 x 2 mL
Subtype:
Cell Isolation Kits
Cell Type:
Cancer Cells and Cell Lines
Species:
Human
Sample Source:
Buffy Coat; Whole Blood
Selection Method:
Negative
Application:
Cell Isolation
Brand:
RosetteSep
Area of Interest:
Cancer Research; Immunology

Scientific Resources

Educational Materials

(4)

Frequently Asked Questions

What is RosetteSep™?

RosetteSep™ is a rapid cell separation procedure for the isolation of purified cells directly from whole blood, without columns or magnets.

How does RosetteSep™ work?

The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a density centrifugation medium (e.g. Ficoll-Paque™ PLUS, Lymphoprep™).

What factors affect cell recovery?

The temperature of the reagents can affect cell recovery. All reagents should be at room temperature (sample, density centrifugation medium, PBS, centrifuge) before performing the isolations. Layering can also affect recovery so be sure to carefully layer the sample to avoid mixing with the density centrifugation medium as much as possible. Be sure to collect the entire enriched culture without disturbing the RBC pellet. A small amount of density centrifugation medium can be collected without worry.

Which cell samples can RosetteSep™ be used with?

RosetteSep™ can be used with leukapheresis samples, bone marrow or buffy coat, as long as: the concentration of cells does not exceed 5 x 107 per mL (can dilute if necessary); and there are at least 100 RBCs for every nucleated cell (RBCs can be added if necessary).

Can RosetteSep™ be used with previously frozen or cultured cells?

Yes. Cells should be re-suspended at 2 - 5 x 107 cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.

Can RosetteSep™ be used to enrich progenitors from cord blood?

Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll™, which can lead to a higher RBC contamination than peripheral blood separations.

Does RosetteSep™ work with mouse cells?

No, but we have developed EasySep™, a magnetic-based cell isolation system which works with mouse and other non-human species.

Which anticoagulant should be used with RosetteSep™?

Peripheral blood should be collected in heparinized Vacutainers. Cord blood should be collected in ACD.

Should the anticoagulant be washed off before using RosetteSep™?

No, the antibody cocktail can be added directly to the sample.
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Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area Workflow Stages for
Workflow Stages

Data and Publications

Data

Typical RosetteSep™ CTC Enrichment Profile

Figure 1. Typical RosetteSep™ CTC Enrichment Profile

In the example above, CAMA (epithelial tumor cell line) cells were seeded into whole blood at a starting frequency of 0.8%. The CAMA cell content of the enriched fraction is 60%. Typically 3.2 to 4.4 log depletion of targeted CD45+ cells is attained.

Publications

(3)
Seminars in cancer biology 2017 APR

How to study and overcome tumor heterogeneity with circulating biomarkers: The breast cancer case.

V. Appierto et al.

Abstract

Breast cancer ranks first among female cancer-related deaths in Western countries. As the primary tumor can often be controlled by surgical resection, the survival of women with breast cancer is closely linked to the incidence of distant metastases. Molecular screening by next generation sequencing highlighted the spatial and temporal heterogeneity of solid tumors as well as the clonal evolution of cancer cells during progression and under treatment pressure. Such findings question whether an optimal assessment of disease progression and a screening for druggable mutations should be based on molecular features of primary or recurrent/metastatic lesions and therefore represent a crucial element for failure or success of personalized medicine. In fact, new targeted therapies may induce only short-term benefit annulled by the emergence of resistant clones with new driver mutations which would need to be rapidly and reliably identified. Serial tissue sampling is therefore essential but, unfortunately, also represents a problem since biopsies from solid lesions, which are invasive and potentially painful and risky, cannot be easily repeatedly sampled, are inaccessible or may not fully reflect tumor heterogeneity. The need to early detect and strike this moving target" is now directing the scientific community towards liquid biopsy-based biomarkers which include circulating tumor cells (CTC) and cell-free circulating tumor DNA (ctDNA) can be repeatedly assessed through non-invasive and easy-to-perform procedures and may act as reliable read-outs of functional and molecular features of recurrent/metastatic lesions. In this review we summarize the outcome of CTCs and ctDNA in breast cancer with special reference on their role on unveiling and overcoming tumor heterogeneity on their potential relevance for tumor surveillance and monitoring and for the selection of therapeutic options. Finally we propose integration between blood-based molecular and clinical approaches for monitoring disease progression according to the specific pattern of recurrence of the most aggressive breast cancer molecular subtypes."
Scientific reports 2016 DEC

Improved detection of circulating tumor cells in non-metastatic high-risk prostate cancer patients.

A. Kuske et al.

Abstract

The relevance of blood-based assays to monitor minimal residual disease (MRD) in non-metastatic prostate cancer (PCa) remains unclear. Proving that clinically relevant circulating tumor cells (CTCs) can be detected with available technologies could address this. This study aimed to improve CTC detection in non-metastatic PCa patients by combining three independent CTC assays: the CellSearch system, an in vivo CellCollector and the EPISPOT. Peripheral blood samples from high-risk PCa patients were screened for CTCs before and three months after radical prostatectomy (RP). Combining the results of both time points, CTCs were detected in 37{\%}, 54.9{\%} and 58.7{\%} of patients using CellSearch, CellCollector and EPISPOT, respectively. The cumulative positivity rate of the three CTC assays was 81.3{\%} (87/107) with 21.5{\%} (23/107) of patients harboring ≥5 CTCs/7.5 ml blood. Matched pair analysis of 30 blood samples taken before and after surgery indicated a significant decrease in CTCs captured by the CellCollector from 66{\%} before RP to 34{\%} after therapy (p = 0.031). CTC detection by EPISPOT before RP significantly correlated with PSA serum values (p {\textless} 0.0001) and clinical tumor stage (p = 0.04), while the other assays showed no significant correlations. In conclusion, CTC-based liquid biopsies have the potential to monitor MRD in patients with non-metastatic prostate cancer.
Nature medicine 2014 AUG

Tumorigenicity and genetic profiling of circulating tumor cells in small-cell lung cancer.

C. L. Hodgkinson et al.

Abstract

Small-cell lung cancer (SCLC), an aggressive neuroendocrine tumor with early dissemination and dismal prognosis, accounts for 15-20{\%} of lung cancer cases and ∼200,000 deaths each year. Most cases are inoperable, and biopsies to investigate SCLC biology are rarely obtainable. Circulating tumor cells (CTCs), which are prevalent in SCLC, present a readily accessible 'liquid biopsy'. Here we show that CTCs from patients with either chemosensitive or chemorefractory SCLC are tumorigenic in immune-compromised mice, and the resultant CTC-derived explants (CDXs) mirror the donor patient's response to platinum and etoposide chemotherapy. Genomic analysis of isolated CTCs revealed considerable similarity to the corresponding CDX. Most marked differences were observed between CDXs from patients with different clinical outcomes. These data demonstrate that CTC molecular analysis via serial blood sampling could facilitate delivery of personalized medicine for SCLC. CDXs are readily passaged, and these unique mouse models provide tractable systems for therapy testing and understanding drug resistance mechanisms.
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.