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Dynabeads™ M-450 Tosylactivated
Invitrogen™

Dynabeads™ M-450 Tosylactivated

Dynabeads™ M-450 Tosylactivated are 4.5 μm superparamagnetic beads containing surface tosyl groups. The beads covalently bind primary amino and sulfhydrylRead more
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Catalog NumberQuantity
140135 mL
Catalog number 14013
Price (USD)
893.65
Online Exclusive
1,224.00
Save 330.35 (27%)
Each
Quantity:
5 mL
Price (USD)
893.65
Online Exclusive
1,224.00
Save 330.35 (27%)
Each
Dynabeads™ M-450 Tosylactivated are 4.5 μm superparamagnetic beads containing surface tosyl groups. The beads covalently bind primary amino and sulfhydryl groups in antibodies to position them in the optimal orientation for cell-surface protein binding. Low background and covalent binding of the antibodies make these beads an excellent choice for cell separation applications when a downstream release from the beads is not required.

• Quickly deplete cells or positively isolate target cells for downstream molecular analysis
• Excellent for cell isolation procedures using your own unique antibody
• Several antibodies can be coupled on the same beads for non-cell-based applications, such as cell activation

Coupling procedure
Covalent coupling is performed overnight by incubating the antibody of choice with the Dynabeads™ M-450 Tosylactivated beads. Optimal coupling occurs at high pH (8.5–9.5) and at 37°C. For pH-labile antibodies, coupling can be performed in an alternative buffer at pH 7.4.

Not recommended for immunoprecipitation (IP)
These 4.5 μm Tosylactivated beads are not recommended for IP as they have a lower surface capacity per unit mass when compared to 1 μm and 2.8 μm Tosylactivated and Epoxy Dynabeads™.

Learn more about Dynabeads™ products
• Find Dynabeads™ products for a whole range of applications.
• Find magnets for Dynabeads™ separations.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Cell TypeAll cells from all species
Isolation TechnologyDepletion, positive isolation for molecular applications
No. of CellsProcesses ∼2 x 109 cells total
Output Viability>95%
Product LineDYNAL, Dynabeads
Purity or Quality GradeResearch Grade
Quantity5 mL
ReactivityAll species
Sample TypePBMC, Tissue Digests, Blood
Shipping ConditionRoom Temperature
Starting Material Cell No.1 x 107 cells per isolation
Target SpeciesAll species
Diameter (Metric)4.5 μm
Product TypeTosylactivated Bead
Unit SizeEach
Contents & Storage
Contains: 5 mL beads at a concentration of 4 x 108 beads/mL in distilled water.
Storage: 2°C to 8°C.

Frequently asked questions (FAQs)

What reactive groups do you offer for your surface-activated Dynabeads magnetic beads?

We offer tosyl, epoxy, carboxylic acid and amine activated Dynabeads magnetic beads. Please see the link (https://tools.thermofisher.com/content/sfs/brochures/Surface_Activated_Dynabeads.PDF) for a comparison of the beads.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What is the density of Dynabeads magnetic beads?

The density of Dynabeads magnetic beads is a challenging property to determine. The reason is that Dynabeads magnetic beads have a 17-37% magnetic iron oxide content in order to have a reasonable magnetic separation time, and the density of the iron oxide is about 4.9 g/cm3. Dynabeads magnetic beads are composite materials, being a mix of polymers and iron oxide, and there are very few polymers that have a density below 1.

The sedimentation rate depends on the bead diameter squared, so the sedimentation of a 1 µm bead is much slower than that of 2.8 µm. The effect of diameter on sedimentation rate is to some extent counteracted by the fact that smaller beads need to have a higher content of iron oxide for magnetic separation applications. Typically, our M-280 Dynabeads (diameter 2.8 µm) have a density of 1.4 g DS/cm3 (DS = dry substance), our M-270 Dynabeads (diameter 2.8 µm) and M-450 Dynabeads (diameter 4.5 µm) have a density of 1.6 g DS/cm3, and our MyOne Dynabeads (diameter 1 µm) have a density of 1.8 g DS/cm3.

Find additional tips, troubleshooting help, and resources within our Dynabeads Nucleic Acid Purification Support Center as well as our Dynabeads Cell Isolation and Expansion Support Center and Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center.

Can I use Dynabeads M-450 magnetic beads for immunoprecipitation (IP)?

Dynabeads M-450 magnetic beads are 4.5 micron beads and may be used for immunoprecipitation. An example is Dynabeads Sheep anti-Rat IgG magnetic beads (Cat. No. 11035) which you may use when your primary antibodies are of rat origin. The smaller beads (2.8 micron) do, however, provide larger surface area and will give higher yields of protein.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Can I store the antibody/ligand-coated Dynabeads magnetic beads during isolation of cell organelles?

Dynabeads magnetic beads coated with antibody/ligand may be stored at 2-8 degrees C for several weeks without loss of antigen binding capacity. After storage, coated Dynabeads magnetic beads should be washed once in PBS/BSA for 5 mins before use.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Which surface-activated Dynabeads magnetic beads should I choose for immobilizing my protein?

This depends on the nature of the specific ligand to be immobilized and the desired downstream application.

-The most frequently used surface-activated Dynabeads magnetic beads for protein isolation are the Tosyl-activated Dynabeads magnetic beads M-280. These beads are hydrophobic, easy to handle and ideal for covalent binding of antibodies for immunoprecipitation. Other ligands could also be covalently bound to these beads, but they have to tolerate conditions like neutral to high pH and high temperatures (required for covalent bond formation).
-Dynabeads magnetic beads M-270 Epoxy are used when the ligand to be immobilized needs to be treated gently and will not tolerate harsh binding conditions like high temperature or pH. Proteins/peptides (other than antibodies) and enzymes are often coupled onto these beads.
-Dynabeads magnetic beads M-270 Amine is often used in combination with crosslinkers to create specific surface groups on the beads. Hetro-bifunctional crosslinkers with an amine-reactive NHS group at one end and another chemical group of your choice at the other end are most frequently used. For example, Dynabeads magnetic beads M-270 Amine can be reacted with a hetero-bifunctional crosslinker containing a NHS group at one end and maleimide at the other end to create Dynabeads magnetic beads with a maleimide surface. Since maleimide reacts specifically with sulfhydryl groups, these modified Dynabeads magnetic beads can be used for applications where binding of sulfhydryl groups are desired. Dynabeads magnetic beads M-270 Amine may also be used for direct ligand coupling via aldehyde or ketone groups by Schiff base (imine) formation and reductive amination. In addition carboxylic acid groups on a ligand can be activated with a carbodiimide like EDC, which results in a direct amide bond formation between the beads and the ligand. Alternatively, a crosslinker may be introduced to the ligand. After activation of the ligand with crosslinker, the free end on the crosslinker has to be amine reactive.
-Dynabeads magnetic beads M-270 Carboxylic Acid and Dynabeads magnetic beads MyOne Carboxylic Acid can also be used for immobilizing proteins. The carboxylic acid groups on these beads need to be activated with a carbodiimide before coupling. The negatively-charged surface of these beads may attract positively charged proteins and cause nonspecific binding. This needs to be considered if these beads are going to be used for immunoprecipitation.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Citations & References (6)

Citations & References
Abstract
Relation of the size and intracellular sorting of apoB to the formation of VLDL 1 and VLDL 2.
Authors:Stillemark-Billton P, Beck C, Borén J, Olofsson SO,
Journal:J Lipid Res
PubMed ID:15520448
'In this study, we tested the hypothesis that two separate pathways, the two-step process and an apolipoprotein B (apoB) size-dependent lipidation process, give rise to different lipoproteins. Expression of apoB-100 and C-terminally truncated forms of apoB-100 in McA-RH7777 cells demonstrated that VLDL particles can be assembled by apoB size-dependent linear ... More
Cell stiffness and receptors: evidence for cytoskeletal subnetworks.
Authors:Huang H, Sylvan J, Jonas M, Barresi R, So PT, Campbell KP, Lee RT,
Journal:Am J Physiol Cell Physiol
PubMed ID:15385268
'Viscoelastic models of cells often treat cells as homogeneous objects. However, studies have demonstrated that cellular properties are local and can change dramatically on the basis of the location probed. Because membrane receptors are linked in various ways to the intracellular space, with some receptors linking to the cytoskeleton and ... More
Isolation and characterization of microvessel endothelial cells from human mammary adipose tissue.
Authors:Hewett PW, Murray JC, Price EA, Watts ME, Woodcock M,
Journal:In Vitro Cell Dev Biol Anim
PubMed ID:7686548
A method for the isolation and long-term culture of human microvessel endothelial cells from mammary adipose tissue (HuMMEC) obtained at breast reduction surgery has been developed. Pure cultures of HuMMEC were isolated by sequential digestion of the fat with collagenase and trypsin followed by specific selection of microvessel fragments with ... More
Human lung microvessel endothelial cells: isolation, culture, and characterization.
Authors:Hewett PW, Murray JC,
Journal:Microvasc Res
PubMed ID:8412855
The pulmonary vasculature is of great physiological/pathological significance. We have isolated and cultured microvessel endothelial cells (HuLEC) from lung tissue obtained from lung transplant recipients by modification of published methods. Pure cultures of HuLEC were isolated by mechanical disaggregation of the tissue prior to sequential dispase and trypsin digestion to ... More
Expansion of functional endogenous antigen-specific CD4+CD25+ regulatory T cells from nonobese diabetic mice.
Authors:Masteller EL, Warner MR, Tang Q, Tarbell KV, McDevitt H, Bluestone JA,
Journal:J Immunol
PubMed ID:16116193
CD4+CD25+Foxp3+ regulatory T cells (T(reg)) are critical for controlling autoimmunity. Evidence suggests that T(reg) development, peripheral maintenance, and suppressive function are dependent on Ag specificity. However, there is little direct evidence that the T(reg) responsible for controlling autoimmunity in NOD mice or other natural settings are Ag specific. In fact, ... More