Dynabeads™ Epithelial Enrich
Dynabeads™ Epithelial Enrich
Invitrogen™

Dynabeads™ Epithelial Enrich

Dynabeads Epithelial Enrich Beads are 4.5-μm superparamagnetic beads coupled to a monoclonal mouse anti-human Ber-EP4 (EpCAM) antibody and intended forRead more
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Catalog number 16102
Price (USD)
1,880.00
Each
Add to cart
Price (USD)
1,880.00
Each
Add to cart
Dynabeads Epithelial Enrich Beads are 4.5-μm superparamagnetic beads coupled to a monoclonal mouse anti-human Ber-EP4 (EpCAM) antibody and intended for isolation of circulating epithelial tumor cells (CTCs) from liquid biopsy samples. The sensitive isolation method provided by Dynabeads Epithelial Enrich Beads is necessary given the low number of CTCs in liquid biopsy samples. The isolated CTCs remain attached to the beads throughout the process.

For downstream molecular or proteomic analysis, the cells can be directly lysed while still attached to the beads. RT-PCR can be performed for amplification and quantification of target nucleic acids. CTC isolation and any subsequent bead-based proteomic or molecular applications can be performed manually or automated using the KingFisher Apex purification system.

Benefits of Dynabeads Epithelial Enrich Beads include:
• High sensitivity and specificity isolation of CTCs from liquid biopsy samples
• Fast and efficient isolation of viable CTCs
• Automated script available for high-throughput enrichment on the KingFisher Apex instrument

Dynabeads-based positive isolation of circulating epithelial tumor cells
Dynabeads Epithelial Enrich Beads are added to the sample under continuous mixing for 30 minutes to optimize the binding of the Dynabeads to the target CTCs. By placing the sample in a DynaMag magnet, the bead-bound epithelial cells are separated from the rest of the sample in < 2 minutes. The supernatant is discarded, and the bead-bound cells are washed 2–3 times to obtain optimal purity. The cells can be lysed while still attached to the beads and the supernatant transferred to a new tube for cell culture or downstream molecular or proteomic analysis. Gene expression analysis without pre-enrichment of RNA can easily be performed using the TaqMan Cells-to-CT Express Kit. Starting samples can be whole blood, buffy coat, bone marrow, or peripheral blood mononuclear cells (PBMCs).

High throughput positive isolation of CTCs on KingFisher Apex instrument
The entire workflow can be automated using the KingFisher Apex purification system with 96-deep-well plates. The high-throughput recovery of bead-bound CTCs can be achieved in ∼50 minutes after the sample and reagents are loaded into the plates. The isolated CTCs are viable and suitable for culture and downstream molecular or proteomic analysis.

Benefits of using automated CTC isolation include:
• Reduced hands-on time
• High reproducibility due to fewer manual handling errors
• Increased throughput with high purity of viable CTCs

Alternative CTC enrichment methods
Positive isolation with release: If it is preferred to positively isolate CTCs for immunological applications such as flow cytometry, we recommend using Dynabeads CELLection Epithelial Enrich Beads (EpCAM, Cat. No. 16203) where the beads are released from the cells after the isolation. The bead-free CTCs are pure, viable and can be used in any downstream application. If you are using your own biotinylated antibody, we recommend using Dynabeads MyOne Streptavidin T1.
Negative isolation of untouched CTCs: If it is preferred to isolate untouched CTCs by leukocyte depletion, we recommend using Dynabeads CD45 Beads (Cat. No. 11153D).

Commercial supply
Our manufacturing sites are ISO 13485-certified. Bead characteristics and manufacturing conditions help ensure consistency and batch reproducibility making them well suited for commercial supply. If you are in the process of customizing Dynabeads Epithelial Enrich Beads in a commercial product or service or need a larger bulk volume of this product, please contact us at oemdynal@lifetech.com or read more on our Dynabeads OEM page.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Cell TypeTumor Cells, Epithelial Cells
Product TypeAntibody Coated Bead
Isolation TechnologyDepletion, positive isolation for molecular applications
No. of CellsProcesses ∼4 x 109 cells total
Output Viability>95%
Product LineDYNAL™, Dynabeads™
Purity or Quality GradeResearch Grade
ReactivityHuman
Sample TypePBMC, Blood
Shipping ConditionRoom Temperature
Starting Material Cell No.2 x 107 PBMCs per isolation
Target SpeciesHuman
Unit SizeEach
Contents & Storage
This Product Contains: 5 mL Dynabeads™ coated with the monoclonal antibody BerEP4 against the human epithelial antigen, EpCAM.

Store at 2°C to 8°C.

Frequently asked questions (FAQs)

My Dynabeads magnetic beads are not pelleting well with the magnet. Do you have any suggestions for me?

Please review the following possibilities for why your Dynabeads magnetic beads are not pelleting:

- The solution is too viscous.
- The beads have formed aggregates because of protein-protein interaction.

Try these suggestions: - Increase separation time (leave tub on magnet for 2-5 minutes)
- Add DNase I to the lysate (~0.01 mg/mL)
- Increase the Tween 20 concentration to ~0.05% of the binding and/or washing buffer.
- Add up to 20 mM beta-merecaptoethanol to the binding and/or wash buffers.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

I have a long double-stranded DNA fragment I would like to isolate. What product do you recommend?

For biotin-labeled DNA that is less than 1 kb, we recommend you use Dynabeads M270 Streptavidin (Cat. No. 65305) and MyOne C1 magnetic beads (Cat. No. 65001). We recommend our Dynabeads KilobaseBINDER Kit (Cat. No. 60101), which is designed to immobilize long (>1 kb) double-stranded DNA molecules. The KilobaseBINDER reagent consists of M-280 Streptavidin-coupled Dynabeads magnetic beads along with a patented immobilization activator in the binding solution to bind to long, biotinylated DNA molecules for isolation. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/immobilisation-of-long-biotinylated-dna-fragments.html) for more information in regards to long biotinylated DNA fragment isolation.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

Can I use Dynabeads magnetic beads to isolate single-stranded DNA templates?

Yes, Dynabeads magnetic beads can be used to isolate single-stranded DNA. Streptavidin Dynabeads magnetic beads can be used to target biotinylated DNA fragments, followed by denaturation of the double-stranded DNA and removal of the non-biotinylated strand. The streptavidin-coupled Dynabeads magnetic beads will not inhibit any enzymatic activity. This enables further handling and manipulation of the bead-bound DNA directly on the solid phase. Please see the following link (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/napamisc/capture-of-biotinylated-targets/preparing-single-stranded-dna-templates.html) for more information in regards to single-stranded DNA capture.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

What is the magnetic susceptibility for Dynabeads magnetic beads?

Magnetic susceptibility is a measure of how quickly the beads will migrate to the magnet. This will depend on the iron content and the character of the iron oxide. The magnetic susceptibility given for the Dynabeads magnetic beads is the mass susceptibility, given either as cgs units/g or m^3/kg (the latter being an SI unit). For ferri- and ferromagnetic substances, the magnetic mass susceptibility is dependent upon the magnetic field strength (H), as the magnetization of such substances is not a linear function of H but approaches a saturation value with increasing field. For that reason, the magnetic mass susceptibility of the Dynabeads magnetic beads is determined by a standardized procedure under fixed conditions. The magnetic mass susceptibility given in our catalog is thus the SI unit. Conversion from Gaussian (cgs, emu) units into SI units for magnetic mass susceptibility is achieved by multiplying the Gaussian factor (emu/g or cgs/g) by 4 pi x 10^-3. The resulting unit is also called the rationalized magnetic mass susceptibility, which should be distinguished from the (SI) dimensionless magnetic susceptibility unit. In general, magnetic mass susceptibility is a measure of the force (Fz) influencing an object positioned in a nonhomogenous magnetic field. The magnetic mass susceptibility of the Dynabeads magnetic beads is measured by weighing a sample, and then subjecting the sample to a magnetic field of known strength. The weight (F1) is then measured, and compared to the weight of the sample when the magnetic field is turned off (F0). The susceptibility is then calculated as K x 10^-3 = [(F1-F0) x m x 0.335 x 10^6], where K is the mass susceptibility of the sample of mass m. The susceptibility is then converted to SI units.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.

How can I determine coupling efficiency of Dynabeads magnetic beads?

There are different methods to check binding of ligands to the beads, including optical density (OD) measurement, fluorescent labeling, and radioactive labeling.

For OD measurement, you would measure the OD of the ligand before immobilization to the beads and compare it with the ligand concentration that is left in the supernatant after coating. This gives a crude measurement of how much protein has bound to the beads.

Protocol:

1.Set spectrophotometer to the right wavelength. As a blank, use the Coupling Buffer.
2.Measure the absorbance of the Pre-Coupling Solution. A further dilution may be necessary to read the absorbance, depending upon the amount of ligand added.
3.Measure the absorbance of the Post-Coupling Solution. A dilution may be necessary to read the absorbance.
4.Calculate the coupling efficiency, expressed as the % protein uptake, as follows. [(Pre-Coupling Solution x D) - (Post-Coupling Solution x D)] x 100/(Pre-Coupling Solution x D) where D = dilution factor.

For fluorescent labeling, we suggest negatively quantifying the amount of ligand bound by measuring ligand remaining in the coupling supernatant (compared to the original sample), rather than directly measuring the ligands on the beads. Add labeled ligand to the beads, and measure how much ligand is left in the supernatant (not bound to the beads). By comparing this with the total amount added in the first place, you can then calculate how much of the ligand that has been bound to the beads. Keep in mind that the Dynabeads magnetic beads are also autofluorescent, which is why direct measuring of fluorescence of the bead-bound ligands is not recommended, but rather this indirect approach. The label could be, for example, FITC/PE. Some researchers perform a direct approach with success (using a flow cytometer).

Radioactive labeling is the most sensitive method of the three, but it is also the most difficult one. It involves radioactively labeling a portion of the ligand. We use radiolabeled I-125 in tracer amounts and mix it with "cold" ligands in a known ratio before coupling. The absolute quantities for the ligand on the beads should be obtained by measuring the beads in a scintillation (gamma) counter and comparing the cpm with a standard.

Protocol:

1.Take out an appropriate amount of beads and wash the beads in 1 mL of binding buffer.
2.Pipette out desired amount of human IgG in a separate tube.
3.Mix the human IgG with I-125-labeled human IgG (30,000 - 100,000 cpm).
4.Dilute the mixture of human IgG and I-125-labeled human IgG to 100 mL in binding buffer.
5.Incubate for 30 minutes at room temperature and measure the cpm in a scintillation counter.
6.Wash the beads (with coating) four times, and measure cpm again.
The % binding is calculated by using the equation : (cpm after washing/cpm before washing)x100%.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center.