Make more informed purchasing decisions with our new product availability and delivery estimate feature, now available on all product pages, in your cart, and during checkout.
Sign In
New to STEMCELL?
Register for an account to quickly and easily purchase products online and for one-click access to all educational content.
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
Request Pricing
Thank you for your interest in this product.
Please provide us with your contact information and your local representative
will contact you with a customized quote. Where appropriate, they can also assist you with a(n):
Estimated delivery time for your area
Product sample or exclusive offer
In-lab demonstration
By submitting this form, you are providing your consent to STEMCELL Technologies Canada Inc. and its subsidiaries and affiliates (“STEMCELL”) to collect and use your information, and send you newsletters and emails in accordance with our privacy policy. Please contact us with any questions that you may have. You can unsubscribe or change your email preferences at any time.
The EasySep™ Human CD3 Positive Selection Kit II is designed to isolate CD3+ cells from fresh or previously frozen peripheral blood mononuclear cells by positive selection. Desired cells are targeted with Tetrameric Antibody Complexes recognizing CD3 and dextran-coated magnetic particles. The cocktail also contains an antibody to human Fc receptor to minimize nonspecific binding. Labeled cells are separated using an EasySep™ magnet without the use of columns. Cells of interest remain in the tube while unwanted cells are poured off. The CD3 antigen is expressed on all T cells and CD56+ NKT cells.
This product replaces the EasySep™ Human CD3 Positive Selection Kit (Catalog #18051) for even faster cell isolations.
Figure 1: Typical FACS Results with EasySep™ Human CD3 Positive Selection Kit II
Starting with a single cell suspension of human PBMCs, the CD3+ cell content of the isolated fraction is typically 99.2 ± 0.2% (mean ± SD), using the purple EasySep™ Magnet.
Figure 2: FACS Data for Anti-Human CD8a Antibody, Clone RPA-T8, FITC-Conjugated
(A) Flow cytometry analysis of human peripheral blood mononuclear cells (PBMCs) labeled with Anti-Human CD8a Antibody, Clone RPA-T8, FITC (Catalog # 60022FI; filled histogram) or a mouse IgG1, kappa FITC isotype control antibody (solid line histogram). (B) Flow cytometry analysis of human PBMCs processed with the EasySep™ Human CD3 Positive Selection Kit (Catalog #17851) and labeled with Anti-Human CD8a Antibody, Clone RPA-T8, FITC. Histograms show labeling of PBMCs (Start) and isolated cells (Isolated). Labeling of start cells with a mouse IgG1, kappa FITC isotype control antibody is shown (solid line histogram).
Figure 3: FACS Data for Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488-Conjugated
(A) Flow cytometry analysis of human peripheral blood mononuclear cells (PBMCs) labeled with Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488 (Catalog #60016AD) and Anti-Human CD45 Antibody, Clone HI30, APC (Catalog #60018AZ). (B) Flow cytometry analysis of human PBMCs isolated with the EasySep™ Human CD3 Positive Selection Kit (Catalog #17851) and labeled with Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488. Histograms show labeling of total PBMCs (Start) and isolated cells (Isolated). Labeling with Mouse IgG2b, kappa Isotype Control Antibody, Clone MPC-11, Alexa Fluor® 488 (Catalog #60072AD) is shown in the bottom panel (solid line histogram). (C) Flow cytometry analysis of human PBMCs isolated with the EasySep™ Human CD4 Positive Selection Kit (Catalog #17852) and labeled with Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488. Histograms show labeling of total PBMCs (Start) and isolated cells (Isolated). Labeling with Mouse IgG2b, kappa Isotype Control Antibody, Clone MPC-11, Alexa Fluor® 488 is shown in the bottom panel (solid line histogram).
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.
Can EasySep™ be used for either positive or negative selection?
Yes. The EasySep™ kits use either a negative selection approach by targeting and removing unwanted cells or a positive selection approach targeting desired cells. Depletion kits are also available for the removal of cells with a specific undesired marker (e.g. GlyA).
How does the separation work?
Magnetic particles are crosslinked to cells using Tetrameric Antibody Complexes (TAC). When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a separate fraction.
Which columns do I use?
The EasySep™ procedure is column-free. That's right - no columns!
How can I analyze the purity of my enriched sample?
The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.
Can EasySep™ separations be automated?
Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.
Can EasySep™ be used to isolate rare cells?
Yes. We recommend a cell concentration of 2x108 cells/mL and a minimum working volume of 100 µL. Samples containing 2x107 cells or fewer should be suspended in 100 µL of buffer.
Are the EasySep™ magnetic particles FACS-compatible?
Yes, the EasySep™ particles are flow cytometry-compatible, as they are very uniform in size and about 5000X smaller than other commercially available magnetic beads used with column-free systems.
Can the EasySep™ magnetic particles be removed after enrichment?
No, but due to the small size of these particles, they will not interfere with downstream applications.
Can I alter the separation time in the magnet?
Yes; however, this may impact the kit's performance. The provided EasySep™ protocols have already been optimized to balance purity, recovery and time spent on the isolation.
For positive selection, can I perform more than 3 separations to increase purity?
Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.
How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?
Hundreds of publications have used cells selected with EasySep™ positive selection kits for functional studies. Our in-house experiments also confirm that selected cells are not functionally altered by the EasySep™ magnetic particles.
If particle binding is a key concern, we offer two options for negative selection. The EasySep™ negative selection kits can isolate untouched cells with comparable purities, while RosetteSep™ can isolate untouched cells directly from whole blood without using particles or magnets.
BiHC, a T-Cell-Engaging Bispecific Recombinant Antibody, Has Potent Cytotoxic Activity Against Her2 Tumor Cells.
Xing J et al.
Translational oncology 2017 OCT
Abstract
Among different cancer immunotherapy approaches, bispecific antibodies (BsAbs) are of great interest due to their ability to recruit immune cells to kill tumor cells directly. Various BsAbs against Her2 tumor cells have been proposed with potent cytotoxic activities. However, most of these formats require extensive processing to obtain heterodimeric bispecific antibodies. In this study, we describe a bispecific antibody, BiHC (bispecific Her2-CD3 antibody), constructed with a single-domain anti-Her2 and a single-chain Fv (variable fragment) of anti-CD3 in an IgG-like format. In contrast to most IgG-like BsAbs, the two arms in BiHC have different molecular weights, making it easier to separate hetero- or homodimers. BiHC can be expressed in Escherichia coli and purified via Protein A affinity chromatography. The purified BiHC can recruit T cells and induce specific cytotoxicity of Her2-expressing tumor cells in vitro. The BiHC can also efficiently inhibit the tumor growth in vivo. Thus, BiHC is a promising candidate for the treatment of Her2-positive cancers.
An optimized protocol for adenosine triphosphate quantification in T lymphocytes of lymphopenic patients.
Girardot T et al.
Journal of immunological methods 2016 OCT
Abstract
In several clinical contexts, the measurement of ATP concentration in T lymphocytes has been proposed as a biomarker of immune status, predictive of secondary infections. However, the use of such biomarker in lymphopenic patients requires some adaptations in the ATP dosage protocol. We used blood from healthy volunteers to determine the optimal experimental settings. We investigated technical aspects such as the type of anticoagulant for blood sampling, the effect of freeze and thaw cycles, the reagent and sample mixing sequence, and the optimal dilution buffer. We also shortened the incubation time to 8h, and even showed that a 30min incubation may be sufficient. To evaluate the ATP rise upon lymphocyte activation, the optimal dose of stimulant was defined to be 4μg/mL of phytohaemagglutinin. Lastly, we determined that the number of T cells needed for this measurement was as low as 50,000, which is compatible with the existing lymphopenia in clinical settings. This optimized protocol appears ready to be assessed in lymphopenic patients to further investigate the interconnection between T lymphocyte metabolism and impaired phenotype and functions.
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
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.