In This Issue


Swine Cytokine Magnetic 7-Plex Panel   Measure Cytokine Proteins in Pig Samples—Swine Cytokine Magnetic 7-Plex Panel
Introducing the Attune® Autosampler   Your Flow Experiments Just Got Faster and Easier—Introducing the Attune® Autosampler
New ABfinity™ Recombinant Oligoclonal Antibodies   ChIP of Histone H3—New ABfinity™ Recombinant Oligoclonal Antibodies
Neurite Outgrowth Staining Kit   Fast Measurement of Neurite Outgrowth and Cell Viability—Neurite Outgrowth Staining Kit
Convenient Packaging for High-Quality Dye Stock Solutions   Molecular Probes® Anhydrous DMSO—Convenient Packaging for High-Quality Dye Stock Solutions
New Conjugates for Flow Cytometry   Stem Cell Antibodies in Multiple Fluorescent Colors—New Conjugates for Flow Cytometry




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BioProbes® Journal for Cell Biology Applications
BioProbes 68  
The Molecular Probes® Handbook
Molecular Probes Handbook


Measure Cytokine Proteins in Pig Samples—Swine Cytokine Magnetic 7-Plex Panel

What It Is
The Swine Cytokine Magnetic 7-Plex Panel for the Luminex® platform is specifically designed for quantifying 7 swine cytokines (IL-1β, IL-4, IL-8, IL-10, IFN-γ, IFN-α, and TNF-α) simultaneously in serum, plasma, and tissue culture supernatant samples. Multiplexing provides more data from each sample, saving both money and time compared to ELISAs.

What It Offers

  • Superior performance—accurate, reproducible, and sensitive quantitation of multiple proteins
  • Magnetic beads—makes washing easier and facilitates the use of automated washing systems
  • Fast and easy protocols—perform your assays and analyze your data typically in less than one day

How It Works
Magnetic microspheres are internally dyed with red and infrared fluorophores of differing intensities. Each bead is assigned a unique bead "region", allowing differentiation of one bead from another. Beads covalently bound to different antibodies can be mixed in the same assay, in a 96-well microplate format. At the completion of the sandwich immunoassay, beads can be read using the Luminex® 200™, FLEXMAP® 3D, or MAGPIX® system and xPONENT® software to distinguish bead color (which identifies the analyte) and to assay signal strength (PE fluorescence intensity).

Swine Cytokine Magnetic 7-Plex Panel assay mechanism   Swine Cytokine Magnetic 7-Plex Panel assay mechanism. A magnetic bead (red) is conjugated to a capture antibody. Through a series of incubation steps, the analyte of interest is sandwiched between the capture and detector antibodies, and a streptavidin-RPE detection system is used for quantification (green).
Product Quantity Cat. No.
Swine Cytokine Magnetic 7-Plex Panel 100 tests LSC0001M

Your Flow Experiments Just Got Faster and Easier—Introducing the Attune® Autosampler

What It Is
The Attune® Autosampler, an optional accessory to the Attune® Acoustic Focusing Cytometer, enables rapid processing of multiple samples. In addition, the newly improved software offers greater data analysis functionality.

What It Offers

  • Wide compatibility—the Attune® Autosampler is compatible with many different formats, including tubes and 96-well, 384-well, and deep-well plates
  • Sample integrity and data quality—the Attune® Autosampler mixes sample by aspiration (instead of shaking) to help ensure sample uniformity
  • Easy operation—the Attune® Autosampler can be easily attached to and detached from the instrument, allowing users to quickly switch between the autosampler and individual tubes

How It Works
The Attune® Acoustic Focusing Cytometer uses proprietary sound wave technology to help ensure accurate alignment of cells through the path of the laser beam. The Attune® Autosampler enables high-throughput sample processing. Its intelligent probe leads to minimal clogging and carryover and also prevents instrument damage. Using the virtual plate layout view, users can create compensation wells, define instrument settings, and identify samples. The free software allows multiple experiments to be defined on a single plate.

The Attune® Acoustic Focusing Cytometer and Attune® Autosampler  
The Attune® Acoustic Focusing Cytometer and Attune® Autosampler.
Product Quantity Cat. No.
Attune® Autosampler with CPU 1 each 4473927
Attune® Autosampler 1 each 4473928

ChIP of Histone H3—New ABfinity™ Recombinant Oligoclonal Antibodies

What They Are
Histone H3 is a member of the nucleosome core around which DNA wraps to form chromatin in eukaryotic cells. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and is subjected to several different types of posttranslational modifications. This histone is known to be the most highly posttranslationally modified of all the five histones and has growing importance in epigenetics and gene regulation. Mammalian cells have three known sequence variants of this protein, called Histone H3.1, Histone H3.2, and Histone H3.3, but the differences comprise just a few amino acids. This antibody specifically recognizes Histone H3 protein with phosphorylated serine at the tenth amino acid position. 

What They Offer

  • Reliability—reproducible lot-to-lot performance
  • Consistency—minimize the need to revalidate working antibody dilutions for your experiments each time you order

How They Work
ABfinity™ antibodies are manufactured by transfecting mammalian cells with high-expression vectors containing immunogen-specific heavy- and light-chain antibody cDNA. ABfinity™ oligoclonal antibodies are a mixture of recombinant monoclonal antibodies, combining the improved signal strength of a polyclonal antibody with the highly reproducible results you get from ABfinity™ monoclonal antibodies.

ChIP qPCR analysis of Histone H3 (pS10) ABfinity™ Recombinant Rabbit Oligoclonal Antibody  
ChIP qPCR analysis of Histone H3 (pS10) ABfinity™ Recombinant Rabbit Oligoclonal Antibody. ChIP was performed on HeLa chromatin using the Histone H3 (pS10) ABfinity™ Recombinant Rabbit Oligoclonal Antibody. Real-time PCR was performed for c-Fos (positive locus) and SAT2 (negative locus) targets using an isotype antibody as a negative control. Data are presented as fold enrichment of the ChIP antibody signal compared to the IgG negative control, determined using the comparative Ct method.

Fast Measurement of Neurite Outgrowth and Cell Viability—Neurite Outgrowth Staining Kit

What It Is
The dual-color Neurite Outgrowth Staining Kit uses a selected combination of fluorescent and background suppression dyes for simple, rapid visualization and relative quantification of neurite outgrowth with cell viability. The assay protocol is just a few steps, allowing data collection in as little as 15 minutes.

What It Offers

  • Speed—stain cells faster than with immunocytochemistry (as little as 15 min)
  • Multiplexing—simultaneously measure neurite outgrowth and cell viability
  • Convenience—use with either live or fixed cells

How It Works
Neurite outgrowth is monitored via bright orange staining of outer cell membrane surfaces. Neuronal cell health is assessed simultaneously using a cell-permeant viability indicator dye that is converted by living cells to emit green fluorescence.

Visualization of primary rat cortex neurons using the Neurite Outgrowth Staining Kit  
Visualization of primary rat cortex neurons using the Neurite Outgrowth Staining Kit. Cryopreserved primary rat cortex neurons were thawed and plated in 96-well format and grown in Neurobasal® medium supplemented with B-27® serum-free supplement for 7 days prior to live-cell staining with the Neurite Outgrowth Staining Kit. The overlay image shows the orange-fluorescent Cell Membrane Stain and green-fluorescent Cell Viability Indicator in the same field of view.
Product Quantity Cat. No.
Neurite Outgrowth Staining Kit
1 kit A15001

Molecular Probes® Anhydrous DMSO—Convenient Packaging for High-Quality Dye Stock Solutions

What It Is
DMSO (dimethyl sulfoxide) is a universal solvent and is required for making stock solutions of most fluorescent dyes and indicators. High-quality anhydrous DMSO is now available from the same trusted source that supplies your fluorescent reagents.

What It Offers

  • Convenience—single-use aliquots
  • Freshness—high-quality DMSO solvent for each stock solution
  • Trusted source—order with your fluorescent dyes

How It Works
Many compounds that are not directly soluble in water can be dissolved with DMSO to create a concentrated stock solution that can be further diluted to the desired final concentration in an aqueous buffer or medium at the time of use. Molecular Probes® DMSO is suitable for preparing stock solutions of many reagents, including amine-reactive probes (e.g., SE- or TFP-reactive esters), thiol-reactive probes (e.g., maleimides), acetoxymethyl (AM) or acetate esters, chemical crosslinkers (e.g., SMCC), and more. The recommended concentrations of these stock solutions are typically between 1 mM and 10 mM.


Product Quantity Cat. No.
DMSO, Anhydrous 10 x 3 mL D12345

Stem Cell Antibodies in Multiple Fluorescent Colors—New Conjugates for Flow Cytometry

What They Are
Antibodies to key stem cell markers, including CD34, Ly-6A/E, and SSEA-4, are now available in multiple fluorescent colors.

What They Offer

  • Flexibility—multiple colors allow you to exploit the full complement of lasers on your instrument
  • Convenience—conjugates are supplied in smaller pack sizes
  • Affordability—our average price is 50% less than other antibodies

How They Work
CD34 is expressed on the majority of hematopoietic stem/progenitor cells and bone marrow stromal cells, and CD34 antibodies are commonly used to gate for stem cells in flow cytometric analyses. The mouse Ly-6A/E antibody is expressed on multipotent hematopoietic stem cells in the bone marrow of mice. Upon activation, the expression of Ly-6A/E (also known as Sca-1) is dramatically upregulated in all strains. The human SSEA-4 (stage-specific embryonic antigen) acts as a surface marker for human embryonic stem cells. Our high-quality antibody conjugates to these stem cell markers have been validated for flow cytometry and offer multiple color choices to fit your instrument and your experiment.

Detection of Ly-6A/E in activated splenocytes  
Detection of Ly-6A/E in activated splenocytes. BALB/c splenocytes were activated by incubation for 48 hr with concanavalin A. After harvesting, the cells were stained with rat anti-mouse Ly-6A/E (Sca-1) FITC conjugate (magenta peak). Cells not treated with concanavalin A do not show expression of Ly-6A/E (unshaded peak).


Rapid Method to Monitor Metabolic Health and Lipid Production in Algae

Microalgae are receiving a lot of attention as a potential source of high-value end products such as pharmaceuticals, nutraceuticals, biofuels, and other biomass co-products. Effective assays to monitor algal production cultures are highly desirable to researchers developing small- and large-scale systems. Using the Tali® Image-Based Cytometer, FDA, and BODIPY® 493/503 stain, we have developed a method that allows researchers to rapidly and quantitatively analyze metabolic activity and lipid production in microalgae samples.


Detection of lipid production using the Tali® Image-Based Cytometer   Detection of lipid production using BODIPY® 493/503 lipid stain and the Tali® Image-Based Cytometer. Chlamydomonas reinhardtii cells were grown in Gibco® TAP medium with nitrogen (A, B) and without nitrogen (C, D), and then stained with BODIPY® 493/503 lipid stain. Samples were analyzed on the Tali® Image-Based Cytometer using the dual fluorescence detection mode (Green + Red). By monitoring fluorescence in the green channel, we found that nitrogen-depleted environments produced algal cells with increased lipids (A vs. C) and decreased chlorophyll (B vs. D).


Detect Signal, Not Background, With Image-iT® FX Signal Enhancer

Image-iT® FX Signal Enhancer is a highly effective product for blocking background staining that arises from nonspecific interactions of a wide variety of fluorescent dyes with cell and tissue constituents. Background staining observed with fluorescent conjugates of streptavidin and antibody conjugates is largely eliminated when Image-iT® FX Signal Enhancer is applied to fixed and permeabilized cells prior to staining. Image-iT® FX Signal Enhancer is supplied in a dropper bottle and is applied directly to slides or coverslips containing cell or tissue samples, prior to immunostaining.

Reducing background when imaging cells. Consult the decision tree for reducing background in cell imaging experiments.
Product Quantity Cat. No.
Image-iT® FX Signal Enhancer
10 mL I36933


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Life in the Lab


Life in the Lab Circular From Life Technologies

Life in the Lab is scientainment for your lab. Check out tips and tricks for lab productivity, along with fun videos, useful downloads, and great product offers across our entire Life Technologies portfolio. Read the latest issue of our Life in the Lab circular at our website, or download your own print copy.

Imaging Corner

Selective Visualization of Artery Walls

Selective labeling and visualization of artery walls in the brain using Alexa Fluor® 633 Hydrazide, Bis(Triethylammonium) Salt. The image is an average-intensity 64-µm z-projection obtained in vivo from the visual cortex of an anesthetized rat using two-photon microscopy. After intravenous injection of Alexa Fluor® 633, arteries become brightly labeled (red vessels) because Alexa Fluor® 633 specifically binds to elastin fibers in artery walls. Endothelial cells in all vessels (arteries, veins, and capillaries) were labeled via a subsequent intravenous injection of green tomato lectin. The green lectin dye is shown as blue in the image so that color-blind readers can distinguish the arteries among other vessel types. In the combined red-blue image, the lectin labeling is not visible in arteries because the red Alexa Fluor® 633 dye is much brighter, masking the weak endothelial labeling. Since Alexa Fluor® 633 does not label veins and capillaries in the brain, the lectin dye (blue) is visible in these non-arteriole vessels. Similar results were obtained in four other mammalian species. This discovery was initially reported in the March 2012 issue ofNature Methods by Dr. Prakash Kara's research team at the Medical University of South Carolina.

From the Bench

Reduced Background Staining Helps Deliver More Conclusive Tissue Modeling Data

Tissue engineering and advanced cell modeling are areas of intense research, with participants across a range of natural science and engineering disciplines. A common requirement is the analysis of how well the models recreate the native tissues they are intended to potentially replace or substitute. High-resolution fluorescence imaging is integral to the analysis of such models, and due to the complexity and multi-dimensionality of these models the imaging challenges are significant. Even limited background signal can obscure critical data. While advances in imaging instrumentation and image analysis algorithms make the acquisition of such high-quality images easier than in the past, background staining challenges remain.

Image-iT® Signal Enhancer has been used by thousands of researchers looking for an easy and effective method to reduce background fluorescence from nonspecifically adsorbing stains and antibody conjugates. Recently the application of this ubiquitous tool has been extended to the field of tissue model imaging. Two recent papers (Desroches et al., Am J Physiol Heart Circ Physiol; and Lasher et al., J Tissue Eng) describe approaches to build and refine complex cell models that structurally and functionally mimic human cardiac tissue. Both research groups analyzed the cell models by high-resolution confocal imaging, in which background was minimized by the use of Image-iT® FX Signal Enhancer. Another study investigated factors that drive cell motility in self-organizing mesenchymal cells in a model for the replacement of endocardium or bone tissue. Increased cell motility results in an unexpected pattern of periodic holes, which characterizes trabecular (spongy) tissue (Chen et al., Interface Focus). In a third example, Keller et al. (PNAS) report an elegant study of the identification and definition of the cell types within the breast that give rise to the two major human breast tumor cell types, thereby identifying their normal precursor cells.

In all four cases, the fluorescence signal-to-noise was enhanced by the use of Image-iT® FX Signal Enhancer, demonstrating the broad utility and flexibility of this unique product to address ubiquitous background fluorescence noise.

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