Dear Researcher,

We are pleased to present the September e-Lynx, a newsletter devoted to keeping our customers up-to-date on new technologies, our new products, suppliers, activities and promotions.

• New Suppliers
  • Isogen Life Science - ProXima Imaging Platform
• New Products
  • Macherey-Nagel – NucleoMag 96 RNA - Magnetic RNA Isolation
  • Macherey-Nagel – NucleoMag 96 Virus - Isolation of Viral DNA/RNA
  • Enzo Life Sciences – FluoForte™ Calcium Assay Kit
• Featured Products
  • Micronic – Sample Tracking Solutions - New Videos
• Promotions
  • NuGEN – 30% OFF Trial offer on the WT-Ovation™ RNA Amplification System for qPCR and Microarray Analysis
  • Elchrom Scientific – Save 20% OFF the Q-EL™330 Digital Recording and Analysis System
  • Active Motif - Save 15% OFF all Products for Epigenetics, Chromatin Biology and Transcriptional Regulation
  • PromoCell - Special Offers on Stem and Blood Cells & Media
  • Greiner Bio-One - 3 Value Added Offers on Labware
  • Polyplus Transfection - Special Pricing on jetPRIME™ DNA and siRNA Transfection Reagent
  • Micronic - SampleWare Software for Sample Data Management for only $130
• Technical Report
  • Transcription Factor NFkB - by Active Motif

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New Suppliers

Isogen Life Science

^MJSBioLynx Inc. now offers the ProXima imaging platform from Isogen Life Science.

ProXima combines innovative technology with high performance and ease of use. By incorporating the latest electronics with patented optics, ProXima meets the requirements of your most demanding applications, even those which normally require the use of expensive laser-scanning imaging systems. ProXima is a fully-upgradeable system, designed to provide you with optimal solutions as your research needs develop and change.

Click here to view the Isogen Life Science ProXima Brochure.

Please contact our Technical Support Team at 1-888-593-5969 / tech@biolynx.ca for additional information.

 

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New Products

Macherey-Nagel – NucleoMag 96 RNA
Magnetic-bead based isolation of RNA from tissue and cell samples

NucleoMag 96 RNA is designed for rapid manual and automated small-scale preparation of highly pure total RNA from tissue or cell samples. Typically, 96 samples can be purified in less than 120 minutes on common automation platforms. The kit provides reagents for the purification of up to 30 µg pure RNA from suitable samples (e.g. 20 mg tissue or 2x10⁶ cells).

• One-tube procedure minimizes risk of cross-contamination


• Small elution volumes ≥ 50 µl possible


• Suitable for manual and automated processing


• Recombinant DNase included


• Reducing agent TCEP included (no b-mercaptoethanol necessary)

Click here to view the NucleoMag 96 RNA user manual.

Product Number	Description	Pkg Size
MCN7443501	NucleoMag 96 RNA	1 x 96 Preps
MCN7443054	NucleoMag 96 RNA	4 x 96 Preps

 

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Macherey-Nagel – NucleoMag 96 Virus
Magnetic-bead based isolation of viral RNA/DNA from serum or plasma for high-throughput in 96-well format.

NucleoMag 96 Virus is designed for rapid manual and automated small-scale preparation of viral RNA / DNA from cell-free body fluids such as serum or plasma. Typically, 96 samples can be purified in less than 120 minutes on common automation platforms. The kit provides reagents for the purification of viral DNA/RNA from up to 200 µl sample volume.

• One-tube procedure minimizes risk of cross-contamination


• Small elution volumes ≥ 50 µl possible


• Yield does not depend on elution volume


• Suitable for manual and automated processing


• Easily adapted to common liquid handling instruments


• Proteinase K and carrier RNA included

Click here to view the NucleoMag 96 Virus user manual

Product Number	Description	Pkg Size
MCN7448001	NucleoMag 96 Virus	1 x 96 Preps
MCN7448004	NucleoMag 96 Virus	4 x 96 Preps

 

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Enzo Life Sciences – FluoForte™ Assay Kit
Brilliant results for Calcium Assay Discovery

The FluoForte™ Assay kit is a homogeneous fluorescence-based assay for detecting intracellular calcium mobilization across a broad spectrum of biological targets.

FluoForte™ dye offers superior measurement of challenging cell lines and receptors. Relative to other commercially available dyes, FluoForte™ dye yields the brightest signal and largest assay window.

 

Click here for more product information

Product Number	Description	Pkg Size
ENZ51017	FluoForte™ Calcium Assay Kit (Starter Pack)	10 x 96 wells
ENZ51016	FluoForte™ Calcium Assay Kit (High-Throughput)	100 x 96 wells
ENZ52014	FluoForte™ Reagent	5 x 50 µg
ENZ52015	FluoForte™ Reagent	1 mg

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Featured Products

Micronic - Sample Tracking Solutions

Micronic has created new videos to demonstrate their equipment and consumable products. Click on the products below to view the video.

• Hamilton Screw Cap DeCapper – Fast and safe screw cap decapping and recapping


• TubeWriter – Automated direct-print labeling for labware


• SampleWare Software – Store, organize and track sample data


• SampleScan Mini – 2D Barcode Reader


• VolumeCheck - Liquid level detection for assay plates or tubes

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Promotions

NuGEN – 30% OFF Trial offer on the WT-Ovation™ RNA Amplification System for qPCR and Microarray Analysis

Perform unlimited real-time PCR assays from clinical samples with NuGEN’s WT-Ovation™ system.

• Expand expression biomarkers to include hundreds of genes, from just 5 ng of total RNA


• Gain long-term access to a faithful representation of the entire transcriptome


• Secure an unlimited supply of valuable clinical samples to share today or save for the future

Until September 30th, 2009, take advantage of NuGEN’s special 30% OFF trial offer.
Click here for details.

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Elchrom Scientific – Save 20% OFF the Q-EL™ 330 Digital Recording and Analysis System

The Q-EL™ 330 Digital Recording and Analysis System is a powerful, flexible and affordable tool providing a complete solution for documentation of electrophoresis gels.

Until November 30th, 2009, Save 20% OFF the Q-EL™ 330 System.

Click here for promotion details.

 

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Active Motif - Save 15% OFF all Active Motif Products.
Products for epigenetics, chromatin biology and transcriptional regulation research.

This offer is valid until December 31st, 2009.
Active Motif products include:

Together, ^MJSBioLynx Inc. and Active Motif can provide you with the tools and support that will help you obtain fast, reliable results.
Click here for promotion details.

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PromoCell - Stem and Blood Cell Promotion

PromoCell offers a collection of adult stem cells from different tissues as well as differentiated blood cells. All cells are obtained from approved donor programs at associated medical centers. High quality media that simulate the in vivo environment as well as optimized differentiation media systems are also available for these cell types.

From August 1st to October 31st, 2009,
Take advantage of the following special offers

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• Buy 3 cases of polypropylene HTS plates, Get 1 case Free
• Buy 1 case of Strip plates, Get 1 case Free
• Buy 2 cases of Tissue Culture dishes, Get a Free case of shorty pipettes

Click here for promotion details.

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Polyplus Transfections - Special Pricing on the jetPRIME™ DNA and siRNA Transfection Reagent

If you routinely perform transient gene over-expression studies and/or gene silencing by RNA interference, the new powerful jetPRIME™ is the reagent for you!

Until September 30th, 2009,
Take advantage of Special Pricing on jetPRIME™
Click here for promotion details

Samples are available!
Contact our Technical Support Team to inquire about obtaining a sample.

Image: GFP expression in HEK-293 cells 24 h after transfection with pCMV-GF plasmid using jetPRIME™.

 

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NFkB is comprised of homo- or heterodimers of different subunits. These subunits are members of the structurally related Rel family of transcription factors. Five different Rel proteins (also called Rel/NFkB proteins) have been identified: p50 (NF-kB1), p52 (NF-kB2), p65 (RelA), RelB and c-Rel⁴. The p50/p65 heterodimer is the major Rel/NFkB complex found in most cells. Rel proteins contain an N-terminal region called the Rel Homology Domain (RHD) that is conserved among the family members and is responsible for DNA-binding and dimerization.

Two groups of Rel/NFkB proteins are typically described: RelA, RelB and c-Rel contain a transactivation domain (TD) in their C-termini, which is required for the transport of active NFkB complexes into the nucleus. In contrast, subunits p50 and p52 do not contain transactivation domains; they are unable to transactivate on their own and must form heterodimers with RelA, RelB or c-Rel. Homodimers of p50 and p52 were even reported to repress kB site-dependent transcription in vivo, possibly by competing with other transcriptionally active dimers (e.g. p50/p65) for DNA binding⁵. NF-kB1 and NF-kB2 are synthesized as the large precursors p105 and p100, respectively. The p105 precursor undergoes constitutive processing to generate the mature subunit p50, while the processing of p100 is tightly regulated to generate the mature subunit p52^{6, 7}.

Inactive NFkB dimers are sequestered in the cytoplasm of cells by the IκB family of inhibitory proteins. IkB proteins are a family of related proteins containing an N-terminal regulatory domain followed by six or more ankyrin repeats. These ankyrin repeats interact with the RHD of NFkB proteins, masking their nuclear localization signal and preventing translocation⁸. The IkB family includes: IkBa, IkBb, IkBe, IkBg and Bcl-3.

Activation of NFkB can occur internally by acetylation of p65 (RelA) usually at lysines 218, 221 or 310. Acetylated NFkB is active and resistant to the inhibitory effects of IkB. Activation of NFkB by external inducers requires the phosphorylation and subsequent degradation of IkB proteins. Phosphorylation of IkB proteins by IkB kinases (IKK) at two serine residues, Ser32 & Ser36, located in the IkB regulatory domain leads to ubiquitination of the inhibitor IkB molecules, which are then degraded by the proteasome. IKK is composed of a heterodimer of catalytic IKKa and IkBb and a regulatory protein NEMO (NFkB Essential Modulator). The activated NFkB complex translocates to the nucleus and binds DNA to regulate gene expression.

In addition to activating genes for inflammation, cell survival and cell proliferation, NFkB also turns on expression of its own repressor, IkBa. The newly synthesized IkBa then re-inhibits NFkB, creating an auto feedback loop. This results in oscillating levels of NFkB activity⁹.

References
1- Gilmore, T.D. (2006) Oncogene 25: 6680-6684.
2- Brasier, A.R. (2006) Cardiovasc. Toxicol 6: 111-130.
3- Perkins, N.D. (2007) Nat. Rev. Mol. Cell Biol. 8: 49-62.
4- Nabel, G.J., Verma, I.M. (1993) Genes Dev. 7: 2063.
5- Lernbecher et al. (1993) Nature 365: 767-770.
6- Karin, M., Ben-Neriah, Y. (2000) Annu. Rev. Immunol. 18: 621-663.
7- Senftleben et al. (2001) Science 293: 1495-1499.
8- Jacobs, M.D., Harrison, S.C. (1998) Cell 95: 749-758.
9- Nelson et al. (2004) Science 306: 704-708.

Active Motif Tools to Analyze NFκB Function

TransAM™ transcription factor DNA-binding ELISAs
Active Motif‘s TransAM Kits are ELISA-based, DNA-binding assays that facilitate the study of transcription factor activation in mammalian tissue and cell culture extracts. The TransAM method provides quantitative results in less than 5 hours, eliminates the use of radioactivity or the need to run gels and is up to 100-fold more sensitive than classic electrophoretic mobility shift assay (EMSA), or gelshift techniques.

TransAM NFκB Kits use a 96-stripwell plate to which a double-stranded oligonucleotide containing the NFkB-binding site has been immobilized. When nuclear or whole-cell extracts are added to the plate, activated NFkB binds to its consensus sequence on the plate-bound oligonucleotide. Next, a primary antibody for either p50, p65, p52, RelB or c-Rel is added. This is detected by incubation with a secondary HRP-conjugated antibody and developing reagent. The colorimetric TransAM format is measured using a spectrophotometer, while the more sensitive chemiluminescent TransAM Kits use a luminometer. The readout provides a quantitative measure of activated NFkB. Recombinant proteins for p65 and p50 are also available separately for use as a protein standard in the TransAM Kits.

Supershift™ and Gelshift™ Kits for traditional EMSAs
While Active Motif believes that its TransAM method is far superior to EMSA, we still offer supershift and gelshift assays to study many different transcription factors, including individual NFkB subunits, or the entire NFkB/Rel Family.

RapidReporter® luciferase reporter assay
Active Motif‘s RapidReporter is a patented luciferase reporter gene assay that provides faster, more pronounced response to stimulation than other luciferase assays. RapidReporter vectors contain destabilizing elements that rapidly degrade both the protein and mRNA. This greatly reduces the background so that small and/or transient effects can be detected.

Vectors are available with an empty Multiple Cloning Site (MCS), or with the NFkB promoter (or other widely studied transcription factor promoters) already cloned into the vector.

LigandLink™ for fluorescent labeling and protein localization studies
LigandLink enables you to label your protein of interest in living cells. Simply express the protein as a LigandLink fusion and add one of the cell permeable LigandLink labels to the medium. Monitor protein localization and protein:protein interactions with either red or green fluorescence.

LigandLink vectors are available with an empty MCS, or with an NFκB p65 fusion. (Other transcription factor fusions are also available.) LigandLink labels include fluorescein or hexachlorofluorescein.

FACE™ cell-based ELISAs
Fast, Activated Cell-based ELISA (FACE) Kits provide a cell-based method to monitor proteins activated by phosphorylation. In this in-cell Western method, cells are cultured in 96-well plates and stimulated to induce the pathway of interest. Following stimulation, the cells are fixed to preserve activation-specific protein modifications, such as phosphorylation. Each well is then incubated with a primary antibody specific for the activated protein of interest, such as total NFkB, phospho-NFkB Ser468, or phospho-NFkB Ser536. Subsequent incubation with secondary HRP-conjugated antibody and developing solution provides a colorimetric or chemiluminescent readout that is quantitative and reproducible. (In addition to NFkB, FACE Kits for over 20 other phospho-proteins are available.

FunctionELISA™ IκBα for analysis of IκBα phosphorylation
Active Motif‘s FunctionELISA Kits offer a rapid method to monitor changes in signal pathway proteins, such as IkBa. FunctionELISA IκBα utilizes the Sandwich ELISA technique to capture and quantifiably measure the amount of phosphorylated IkBa present in a sample. This kit has the ability to assay both cell and tissue samples without the need to run, blot and develop gels. (FunctionELISA Kits for TRAIL and cytochrome c are also available.)

ChIP-IT™ Express for binding site analysis
Active Motif offers ChIP-IT Express Kits for chromatin immunoprecipitation (ChIP) to study transcription factor binding sites. ChIP-IT Express utilizes magnetic protein G beads, which is a large improvement over agarose bead-based ChIP, and is available in either a sonication or enzymatic format for chromatin preparation. Active Motif also offers a large number of ChIP-validated antibodies and Ready-to-ChIP Chromatin, which help you perform ChIP successfully.

Universal Magnetic Co-IP to detect protein interactions
Active Motif‘s Universal Magnetic Co-IP Kit enables co-immunoprecipitation for both nuclear and whole-cell complexes. It takes advantage of the same magnetic bead-based protocol utilized in ChIP-IT Express to speed and simplify the IP and wash steps, while greatly reducing background.

SUMOlink™ for SUMOylation studies
Active Motif‘s SUMOlink Kits are used to study SUMOylation, a post-translational modification that affects many biological processes. SUMO is a small ubiquitin-like modifier that shares different degrees of sequence identity with ubiquitin. SUMOlink provides a simple method for generating and detecting SUMOylated proteins in vitro to study either SUMO-1 or SUMO-2 & SUMO-3 activity. The kits provide all the reagents required for the SUMOylation including positive and negative controls to help ensure success.

Recombinant proteins
Active Motif carries recombinant proteins for many transcription factors and cell signaling-related proteins, including NFκB p65 and NFκB p50 proteins that have been validated for use as protein standards in TransAM Transcription Factor ELISAs.

Antibodies
Active Motif‘s offers a broad line of mono- and polyclonal antibodies directed against Rel/NFkB, IkB and IKK proteins among others. Active Motif‘s antibodies are highly characterized, and many have been validated for use in ChIP, Western blot or EMSA.

Cell extracts
Active Motif offers high-quality nuclear, cytoplasmic and whole-cell extracts from a variety of cell types and tissue sources. Many of our extracts are prepared from cells that have been treated to specifically induce activation of hard-to-detect transcription factors. Our ready-to-use extracts are ideal as positive controls in a variety of applications, including TransAM, gelshift and Western blots.

Active Motif has a wide selection of extracts to choose from, many of which are ideal controls for NFkB studies.

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