Click-iT Plus EdU Assay Kits: Now with Alexa Fluor 350 and Alexa Fluor 594 dyes

To expand the multiplexability of the Invitrogen™ Click-iT™ Plus EdU assay for flow cytometry, we have recently introduced kits with either Alexa Fluor™ 350 or Alexa Fluor™ 594 dye, which allow detection of the EdU signal using a UV or 532/561 nm laser, respectively. These two kits join our family of Click-iT Plus EdU Flow Cytometry Assay Kits (see product list), providing maximum flexibility when designing an EdU-based cell proliferation experiment.

The Click-iT EdU cell proliferation assay utilizes click chemistry to provide a superior alternative to traditional BrdU methods for labeling newly synthesized DNA. When the modified nucleoside EdU (5-ethynyl-2´-deoxyuridine) is incorporated during DNA synthesis, it can be detected by a quick click reaction, with minimal disruption to the cell. Unlike antibody-based BrdU assays, Click-iT Plus EdU assays employ a small fluorescent picolyl azide for detection of the incorporated EdU and thus do not require harsh DNA denaturation treatments. Furthermore, these fluorescence-based Click-iT Plus EdU assays can be multiplexed with fluorescent proteins such as R-PE, R-PE tandems, and GFP. The Alexa Fluor 350 kit can be multiplexed with other violet-excitable dyes such as Pacific Orange™ conjugates, which emit at ~550 nm.

 

Click-iT Plus EdU Assay Kits

  Detection of cell proliferation with Click-iT Plus EdU Flow Cytometry Assay Kits. Jurkat (human T-cell leukemia) cells were treated with 10 μM EdU and detected according to the recommended Click-iT™ Plus EdU staining protocol. The figures each show a clear separation of nonproliferating cells and proliferating cells, which have incorporated EdU and been labeled with either (A) Alexa Fluor™ 350 picolyl azide or (B) Alexa Fluor™ 594 picolyl azide.

Human Adipokine 14-Plex Magnetic Panel for the Luminex platform

Adipokines—diverse peptides secreted by adipose (fat) tissue—are a group of cytokines, hormones, and other signaling proteins that play an important role in many health problems, including obesity, diabetes, metabolic disorders, and immune-related diseases. The Invitrogen™ Human Adipokine 14-Plex Magnetic Panel for the Luminex™ platform enables researchers to quickly measure multiple proteins in one well in order to achieve a more holistic understanding of cell signaling interactions. Requiring only 25 μL of serum per well, this Luminex assay panel interrogates 14 targets in a single day. Not only is multiplexing efficient, it also reduces the variability of running assays on different days and samples. This panel is suitable for use with the Luminex™ 200™, FLEXMAP 3D™, and MAGPIX™ systems.

 

Human Adipokine 14-Plex Magnetic Panel for the Luminex platform  
Measurement of serum levels of 14 protein targets in one assay run. Sera from normal and diseased patients were tested with the Human Adipokine 14-Plex Magnetic Panel using the Luminex™ 200™ System.

Alexa Fluor conjugates now available for epitope-tag and loading-control antibodies

We offer a wide range of epitope-tag and loading-control antibodies conjugated with Invitrogen™ Alexa Fluor™ dyes, including green-fluorescent Alexa Fluor 488, orange-fluorescent Alexa Fluor 555, and far red–fluorescent Alexa Fluor 647 dyes. The epitope-tag antibodies are highly specific monoclonal and polyclonal antibodies that recognize commonly used epitopes, including FLAG, GST, HA, His, Myc, and V5. The loading-control antibodies recognize the most common loading and expression control proteins, including GAPDH, β-actin, and β-tubulin. They are used to normalize signals on western blots in order to distinguish expression level differences from loading variability in the samples, and also as complementary stains in immunofluorescence studies. In addition to the Alexa Fluor conjugates, the epitope-tag and loading-control antibodies are available unconjugated or conjugated to biotin, horseradish peroxidase (HRP), or Thermo Scientific™ DyLight™ dyes.

Alexa Fluor conjugates for epitope-tag and loading-control antibodies

 

 

 

Western blot analysis of β-actin and β-tubulin in five whole-cell lysates. Proteins transferred to a Low- Fluorescence PVDF Membrane were probed with the Alexa Fluor™ 488 β-Actin and Alexa Fluor™ 555 β-Tubulin Loading Control Antibodies.


New interleukin primary antibody conjugates for flow cytometry

Antibodies validated for use in flow cytometry provide researchers with the ability to both identify specific cell types and analyze the functional responses of heterogeneous cell populations. Part of the cytokine family, interleukins are intracellular proteins produced by a variety of cell types and used to study cell signaling. Our anti-interleukin antibodies now include brightly fluorescent fluorescein, R-PE, and APC conjugates of anti–IL-27 antibodies for mouse and human cells.

Specificity of a fluorescent anti–IL-27 conjugate  
Specificity of a fluorescent anti–IL-27 conjugate. Human peripheral blood mononuclear cells (PBMCs) were stained with R-PE mouse anti–human IL-27 antibody (orange) or isotype-control antibody (blue) and analyzed by flow cytometry.

ArC kits for compensation of LIVE/DEAD Fixable Dead Cell Stains

The Invitrogen™ ArC™ Amine-Reactive Compensation Bead Kit, now available in two sizes, is a bead-based compensation tool specifically optimized for use with the Invitrogen™ LIVE/DEAD™ Fixable Dead Cell Stain Kits but also compatible with other assays that use amine-reactive dyes. Established as a ready-to-use control for setting compensation on flow cytometers, the ArC amine-reactive beads eliminate the hassle of heat-treating cells as a control, thus saving your experimental samples while enabling accurate and consistent results.

Use of ArC beads to mimic staining with LIVE/ DEAD Fixable Dead Cell Stains  
Use of ArC beads to mimic staining with LIVE/ DEAD Fixable Dead Cell Stains. A mixture of positive and negative beads from the ArC™ Amine-Reactive Compensation Bead Kit was stained with LIVE/DEAD™ Fixable Violet Dead Cell Stain, washed, and analyzed by flow cytometry using a 405 nm laser with 450/50 nm bandpass filter.