Fluo-4 now in a complete kit for calcium imaging applications

Fluo-4 Calcium Imaging Kit

The Fluo-4 Calcium Imaging Kit has been designed for the specific detection of calcium flux in imaging applications. Upon binding calcium, the fluo-4 dye exhibits a large fluorescence emission increase (>100-fold) with a minimal shift in wavelength maximum; the emission from calcium-bound fluo-4 dye (excitation/emission maxima ~494/506 nm) can be detected using standard fluorescein (FITC) filters.

To save you time, we have collected all the reagents you need for detecting calcium flux in live cells with the fluo-4 dye and packaged them into the Fluo-4 Calcium Imaging Kit. In addition to fluo-4 AM, which is supplied as a 1,000X solution in DMSO, this kit contains 100X PowerLoad™ Concentrate for easy cell loading, a 10X Neuro Background Suppressor, and probenecid. The Fluo-4 Calcium Imaging Kit has been formulated, optimized, and validated for imaging applications.

Fluo-4 Calcium Imaging Kit  

 

Calcium ion flux visualized. Cytosolic calcium flux from neuronal cells detected using the Fluo-4 Calcium Imaging Kit.

 
Catalog # Name Size List Price (USD) Qty
F10489 Fluo-4 Calcium Imaging Kit 1 kit 400.00

The next generation of membrane potential probes has arrived

FluoVolt™ Membrane Potential Kit

The FluoVolt™ membrane potential dye represents the next generation in voltage-sensitive fluorescent probes, displaying the best characteristics of the fast- and slow-response membrane potential probes. The FluoVolt™ probe not only responds to changes in membrane potential in less than a millisecond like the fast-response probes, but also displays a high-magnitude fluorescence response like the slow-response probes.

The FluoVolt™ membrane potential dye is a fast-response probe with a superior potential-dependent fluorescence response. The response is fast enough to detect transient (millisecond) potential changes in excitable cells, and the probe generates a signal change in excess of 25% per 100 mV. Compatible with standard fluorescein (FITC) settings, the FluoVolt™ membrane potential dye can be used for imaging electrical activity of intact heart tissues, mapping membrane potentials along neurons and muscle fibers, or measuring potential changes in response to pharmacological stimuli.

Membrane voltage changes measured using the FluoVolt™ Membrane Potential Kit. In panels A and B, differentiated NG-108 cells (mouse neuroblastoma–rat glioma hybrid) were loaded with the FluoVolt™ membrane potential dye. (A) Cells were imaged with 10 msec illumination pulses and images acquired with 2x binning. The three selected traces (B) show fluorogenic responses from the dye as the selected cells (numbered dots in A) spontaneously depolarized and repolarized in culture. For the traces in panels C and D, human HEK 293 (embryonic kidney) cells were loaded with the FluoVolt™ membrane potential dye, imaged with 10 msec illumination pulses, and data acquired with 2x binning. Traces show fluorogenic responses as cells are depolarized at 2 sec intervals from –100 mV to +30 mV (C) or in single steps from –80 mV to 0 mV at 2 sec intervals (D) .

Catalog # Name Size List Price (USD) Qty
F10488 FluoVolt™ Membrane Potential Kit 1 kit 425.00

Express almost any gene using BacMam technology

ViraPower™ BacMam Expression System

The ViraPower™ BacMam Expression System provides a convenient means of expressing almost any gene of interest in mammalian cells with the use of BacMam technology. The BacMam system allows expression of genes up to 38 kb, making it possible to clone and express most genes with introns and then let the cell do the RNA processing and posttranslational modification. Among all the methods available for transfection, the BacMam gene delivery and expression system produces the least cellular toxicity. And because expression is transient and baculoviruses do not replicate in mammalian cells, special containment measures are not required.

The BacMam pCMV-Dest Vector, included as part of the ViraPower™ BacMam Expression System, combines Gateway® cloning and BacMam gene expression technologies for easy recombination-based cloning and baculovirus-based expression of a target gene in variety of cell types. In BacMam technology, a modified insect cell virus (baculovirus) is used as a vehicle to efficiently deliver and transiently express genes in mammalian cells with minimum effort and toxicity. This technology allows for control over the level of expression through the increase or decrease of viral particle concentration. When BacMam technology is combined with Gateway® cloning technology, a variety of sizes of target genes can be cloned and expressed. Sizes ranging from an insert for RNAi to a large gene with introns (38 kb) have been cloned and expressed successfully using the BacMam pCMV-Dest Vector. This plasmid accommodates various cloning schemes, including Gateway®, GeneArt® Seamless Cloning and Assembly, and traditional cloning using restriction enzymes.

BacMam technology  

 

BacMam-mediated gene delivery. BacMam particles are taken up by endocytosis and their contents released for transcription and expression (following migration to the nucleus). Gene expression begins within 4 to 6 hr of transduction and nears its maximum level within 24 hr of transduction.

Catalog # Name Size List Price (USD) Qty
A24227 ViraPower™ BacMam Expression System 1 kit 1,100.00
A24223 BacMam pCMV-Dest Vector 10 µg 598.00

Isolate intact exosomes from a variety of starting samples

Total Exosome Isolation Reagents

With a variety of starting samples, you can now easily enrich for intact exosomes using a method that is much easier and less tedious than ultracentrifugation. We provide five Total Exosome Isolation Reagents, each optimized for enriching exosomes from cell culture media, serum, plasma, urine, or other body fluids. Their simple and reliable protocols can be scaled to a range of sample sizes and typically require only 15–20 minutes of hands-on time.

The flexible and scalable Total Exosome Isolation Reagents allow for fast and efficient exosome enrichment. After your sample is incubated with the Total Exosome Isolation Reagent overnight at 2–8°C, exosomes can be recovered using a standard benchtop centrifuge to spin them at 10,000 x g for 60 minutes. Simply resuspend the pellet, and the exosomes are ready for downstream analysis.

Exosomes    
     
Analysis of exosomes recovered from HeLa cell media. (A) Exosomes recovered with Total Exosome Isolation Reagent (from cell culture media) have a size distribution comparable to (B) exosomes isolated following a traditional sucrose gradient ultracentrifugation protocol. Profiles analyzed on a NanoSight® LM10 instrument show all particles to be smaller than 300 nm; most are about 50–150 nm in size.    
For Research Use Only. Not for use in diagnostic procedures.