e-Lynx - October 2008Dear Researcher, We are pleased to present the October e-Lynx, a newsletter devoted to keeping our customers up-to -date on new technologies, our new products, suppliers, activities and promotions.
New ProductsAssay Designs - MultiBead™
Multiplex Immunoassays for Flow Cytometry
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| Product Number | Description | Pkg Size |
| ADI980001 | Inflammation 8-Plex Panel Pre-mixed bead panel for simultaneous quantification of IFN-gamma, IL-1beta, IL-4, IL-6, IL-8, PGE2, TNF-alpha, and TXB2 via flow cytometry |
1 x 96 wells |
| ADI985002 | IFN-gamma Singleplex Kit Singleplex bead kit for quantification of human IFN-gamma via flow cytometry |
1 x 96 wells |
| ADI985003 | IL-1beta Singleplex Kit |
1 x 96 wells |
| ADI985006 | IL-4 Singleplex Kit Singleplex bead kit for quantification of human IL-4 via flow cytometry |
1 x 96 wells |
| ADI985005 | IL-6 Singleplex Kit Singleplex bead kit for quantification of human IL-6 via flow cytometry |
1 x 96 wells |
| ADI985004 | IL-8 Singleplex Kit Singleplex bead kit for quantification of human IL-8 via flow cytometry |
1 x 96 wells |
| ADI985008 | PGE2 Singleplex Kit Singleplex bead kit for quantification of PGE2 via flow cytometry |
1 x 96 wells |
| ADI985001 | TNF-alpha Singleplex Kit Singleplex bead kit for quantification of human TNF-alpha via flow cytometry. |
1 x 96 wells |
| ADI985007 | TXB2 Singleplex Kit Singleplex bead kit for quantification of TXB2 via flow cytometry. |
1 x 96 wells |
| ADI987001 | Extracellular Buffer Pack I Buffer pack intended for use with IFN-gamma, IL-1beta, IL-4, IL-6, IL-8, PGE2, TNF-alpha, and TXB2 singleplex bead kits. |
1 x 96 wells |
| ADI280001 | MultiBead Analysis Software Generate standard curves and determine sample concentrations using FCS or LMD data files |
1 CD |
Custom MultiBead™ Assay Services are also available. Please contact our Technical Support Department for more information
Assay
Designs - ImmunoSet™ ELISA Development Reagents
An economical way to get the most
out of your research budget
ImmunoSet products for intracellular analytes contain the basic components and protocol for the development of five 96 well colorimetric enzyme immunoassays. They are designed to work with samples diluted to a final volume of 100 µL/well. Each ImmunoSet contains: capture antibody, biotinylated detection antibody, standard and streptavidin conjugated to horseradish peroxidase.
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Sensitivity – the limit of detection is determined using data from multiple assays
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Specificity – potential reactivity to related molecules and species is evaluated
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Dilutional linearity – the minimum recommended dilution to eliminate matrix interference in the assay is determined
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Parallelism – we ensure the recombinant standard used in the assay reflects the native analyte in biological samples
| Product Number | Description | Pkg Size |
| ADI960810 | HO-1 (rat) ImmunoSet | 5 x 96 wells |
| ADI960076 | HO-1 (mouse) ImmunoSet | 5 x 96 wells |
| Coming Soon | p53/MDM2 Complex ImmunoSet |
Active
Motif - Re-ChIP-IT™ for Sequential Immunoprecipitation
Localize two different proteins to
the same genomic locus
When performing chromatin immunoprecipitation (ChIP) experiments, it is often useful to prove that two different proteins or histone modifications are present at the same site in the genome. Or, you may want to determine if a protein coincides with a specific histone modification at the same regulatory element. Re-ChIP (aka Sequential ChIP, Chromatin Re-IP and ChIP Re-ChIP) is a relatively new technique that enables sequential chromatin immunoprecipitations to be performed using two different antibodies so that you can assay for the simultaneous presence of two proteins or distinct histone modifications at the same genomic region of interest.
A complete solution for rapid and efficient Re-ChIP.
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Convenient magnetic bead-based protocol
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Fewer cells required than other Re-ChIP methods
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Compatible with multi-channel pipetting
Sequential Chromatin Immunoprecipitation using Re-ChIP-IT - The lane numbers are the same in each panel to indicate that the DNA is from the same chromatin sample. The left panel shows the results of PCR performed on an aliquot of DNA removed from the experiment after the first ChIP step; the right panel represents PCR results on DNA from chromatin samples after both ChIP steps. For example, chromatin samples subjected to first ChIP using Mouse IgG as a negative control (lanes 1 and 2 in the left panel) were then subjected to a second ChIP with an RNA Pol II antibody (lanes 1 and 2 in the right panel). Chromatin samples in which Mouse IgG was used as either the first antibody (lanes 1 and 2) or second antibody (lanes 5 and 6) show little amplification of GAPDH DNA in either the left (first ChIP) or right panel (first and second ChIP). Chromatin samples in which the first antibody used was anti-RNA Pol II and the second antibody was anti-TFIIB (lanes 3 and 4) show good amplification of GAPDH DNA after the second ChIP (right panel) indicating the co-localization of RNA Pol II and TFIIB at the same region of the GAPDH promoter.
| Product Number | Description | Pkg Size |
| AM53016 | Re-ChIP-IT™ | 25 rxns |
New Literature
Macherey-Nagel
- 2009 Bioanalysis Catalogue
This new catalogue features Macherey-Nagel's line of ready-to-use kits and consumables for purification of nucleic acids and proteins. The products cover a range of applications, providing one-stop-shopping.
Click here to request a copy!
Promotions
Micronic - Incredible Savings on Products for Sample Storage and Traceability
Micronic
offers a unique product line that ensures sample
integrity and traceability. For a limited time,
take advantage of special pricing on selected products
including Traxis and Tracker starter packs.
Click here for promotion details.
USB
- Save 20% OFF USB Molecular Biology Ladders
For a limited time, save 20% OFF USB's Ladders
for DNA, RNA and Proteins.
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here for promotion details.
USB
- Spend $150 on USB Biochemicals and
we'll send you the USB fitness pack Free!
USB
provides biochemicals that deliver reliable and
consistent results. USB biochemicals and Ultrapure
brand products meet the highest industry standards
for quality. They are available in a variety of
grades such as ACS Reagent Grade, CP Grade (Chemically
Pure), USP Grade (USBioAnalyzed) and Molecular Biology
Grade.
Don't forget about their line of convenience reagents. USB offers an extensive selection of Ultrapure biochemical reagents in premixed, ready-to-use formats for convenience in any lab.
Now's a great time to step forward and purchase USB Biochemicals and convenience reagents. When you spend $150 or more, we'll send you a USB water bottle and pedometer to track your own fitness results.
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Active Motif - Order ChIP-IT™ Express HT and Receive a FREE MAG-96 Magnetic Stand
ChIP-IT™ Express HT allows you to perform chromatin IP in a fast, reproducible high-throughput format. It combines the efficiency of the groundbreaking magnetic bead-based ChIP-IT™ Express Kit with a 96-well plate format, enabling the rapid and efficient processing of a large number of ChIP reactions.
Click here for promotion details
Simport
- 25% OFF LockMailer™ Microscope Slide Jar
Get a 25% Discount
on Simports new LockMailer™ Jars,
tamperproof multi purpose containers for mailing,
staining or storing microscope slides.
Click here for promotion details
Assay
Designs - Buy 3, Get 1 Free - Antibody and Protein
Promotion
Offer extended until December 31st, 2008
Fall into savings with this buy 3, get 1 free promotion on Assay Designs top selling antibodies and proteins. All you need to do is select at least 4 Assay Designs™ antibodies or proteins from the list below and you will receive the lowest priced product free of charge.
Click here for promotion details
Technical Report
Fast and Reliable HLA
Typing on Precast PCR-CheckIT Gels
by Elchrom Scientific
Introduction
The Human Leukocyte Antigen System (HLA) is part of the genetic region of the Major Histocompatibility Complex of humans. Proteins expressed by HLA are presented at the surface of cells. HLA proteins are highly diverse and unique to each person and allow the immune system to distinguish between self- and non-self cells. HLA plays a key role in the defence against pathogens, disease defence, predisposition to diseases, drug sensitivity, reproduction, cancer, autoimmunity.
The vital role of the HLA in the immune function has implications when transplanting organs. A close match between the HLAs of donor and recipient is imperative to promote grafting and to avoid post-transplant complications such as graft-versus-host-disease (e.g. Kaneku; Luque; Mao).
Transplantation success is highly correlated with the matching of six HLA markers. Testing HLA markers through PCR-based techniques is, thus, vital for tissue matching (Robinson; Zetterquist). Since 70% of the patients receiving transplants have no suitable donor in their family, diagnostic HLA testing is key to safeguard a transplantation by finding a perfectly matched donor (National Bone Marrow Program). Finding an HLA-compatible donor is a key to allogeneic transplantation of hematopoietic cells to treat hematologic malignancies, too (Demirer).
SSO and SSP Method
There are two methods of choice for HLA kits:Sequence Specific Oligonucleotides (SSO) and Sequence Specific Primers (SSP). Both allowamplification of polymorphic target sequences in the HLA locus by the Polymerase chain reaction. Polymorphic HLA genes (such as e.g. HLA-A, B, C, DP, DQ, and DR) can be routinely typed for in HLA laboratories (Marsh et al; Zetterquist). If a cost-effective system is at hand, large populations of normal and affected individuals can be screened. Analysis of diagnostic bands through electrophoresis then allows matching transplant donors and recipients (Wordsworth; Erlich). Elchrom's Standard Operating Procedure defined PCR-CheckIT Wide Mini S-4x25 gels to be the most effective format to analyze a typical standard set of HLA markers per patient, amplified in 96-well plates.
These gel run in ORIGINS by ElchromTM electrophoresis system allows for:
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Fast results - PCR analysis in as little as 7 minutes
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Most economical solution - PCR-CheckIT gelscan be reused up to 10 times
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More cost and time effective - compared toordinary agarose gels
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Simple handling and maintenance -compared to capillary electrophoresis
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Easy handling - precast PCR CheckIT gelsare mechanically and temperature stable
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Highly reliable system - delivers results ofthe highest fragment size accuracy
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Stable gels - shelf-life of 18 months
Fig 1-6 - Two PCR CheckIT Wide
Mini S-4x25 gels were run and reused for 6 times. Different
sets of HLA samples were run. Only a small sections of the
two gels (9 sample wells), with 8 samples and a size marker,
are shown. Size marker contains 3 DNA fragments of 200, 500
and 1000.
Results
Samples were analysed by electrophoresis on PCR CheckIT gels (Fig. 1-6). The PCR products were directly loaded on a gel using 8-channel electronic equalizer pipette and separated on PCR CheckIT Wide Mini S-4x25 gels with EtBr. After the analysis gels were cleared after each run and reused up to 10 times. Accurate identification of PCR products was easily done after each run by standard HLA analysis software.
Experimental Procedure
Equipment
Electrophoresis was performed in ORIGINS by ElchromTM submarine
electrophoresis apparatus, at 12V/cm (144V). The temperature
of the running buffer was kept constant at 20°C, using
the internal temperature control system.
Sample Preparation and Running
Conditions HLA fragments were amplified by PCR as described
in User Manual of commercially available kits. 8 µl
of sample was loaded onto the PCR CheckIT Wide Mini S-4x25
gel with EtBr. Electrophoresis was performed for 7 min at
144V and 20°C in 30 mM TAE running buffer containing EtBr
at concentration of 0.5 µg/ml. Prior to reuse the gel
was electrophoretically cleaned. To clean the gel DNA was
run out by reversing the gel 180° in the electrophoresis
apparatus and run for 20 minutes at 180V and 20°C.
Detection
Elchrom’s PCR CheckIT Wide Mini S-4x25 were prestained
with Ethidium Bromide and the results were visible immediately
after electrophoresis by viewing the gel on UV transilluminator.
References
Aribi M. Candidate genes implicated in type 1 diabetes susceptibility.
Curr Diabetes Rev. 2008 May;4(2):110-21.
Demirer T, Barkholt L, Blaise D, Pedrazzoli P, Aglietta M,
Carella AM, Bay JO, Arpaci F, Rosti G, Gurman G, Niederwieser
D, Bregni M; EBMT Solid Tumors Working Party. Transplantation
of allogeneic hematopoietic stem cells: an emerging treatment
modality for solid tumors. Nat Clin Pract Oncol. 2008 May;5(5):256-67.
Erlich H, Bugawan T, Begovich A, Scharf S. Analysis of HLA
class II polymorphism using polymerase chain reaction. Arch
Pathol Lab Med. 1993 May;117(5):482-5.
Kaneku H. Detection of soluble HLA-G and its correlation with
kidney transplant outcome. Clin Transpl. 2006:447-54.
Luque J, Torres MI, Aumente MD, Marín J, García-Jurado
G, González R, Pascual D, Guerra N, López-Rubio
F, Alvarez-López MR, Arizón JM, Peña
J. Soluble HLA-G in heart transplantation: their relationship
to rejection episodes and immunosuppressive therapy. Hum Immunol.
2006 Apr-May;67(4-5):257-63.
Mao Q, Terasaki PI, Cai J, Briley K, Catrou P, Haisch C, Rebellato
L. Extremely high association between appearance of HLA antibodies
and failure of kidney grafts in a five-year longitudinal study.
Am J Transplant. 2007 Apr;7(4):864-71.
Marsh S, Parham P, Barber L. The HLA Factsbook. Academic Press
Nagorsen D, Thiel E. HLA typing demands for peptide-based
anti-cancer vaccine. Cancer Immunol Immunother. 2008 Mar 4.
National Marrow Donor Program. HLA Matching: Finding the Best
Donor or Cord Blood Unit. http://www.marrow.org/PATIENT/Donor_
Select_Tx_Process/The_Search_Process/HLA_Matching_Finding_the_Best_/index.html#basics
Robinson J, Marsh SG. The IMGT/HLA database. Methods Mol Biol.
2007;409:43-60.
Wordsworth P. PCR-SSO typing in HLA-diseaseassociation studies.
Eur J Immunogenet.1991 Feb-Apr;18(1-2):139-46.
Zetterquist H, Bengtsson M, Bäckström G,Egle-Jansson
I, Ekdahl AM, Grunnet N,Gustafsson I, Knutsen I, Kuhle A,
RydbergL, Spurkland A, Steffensen R, StorgärdsM, Szojmer
E, Söderholm G, Thuresson B,Turesson H, Olerup O. Report
from the HLAclass II typing by PCR-SSP Multicentre Study.
Eur J Immunogenet. 1997 Jun;24(3):191-9.
Announcement
MJSBioLynx
Inc - Technical Sales Representative
Central and SouthWestern Ontario
We are pleased to announce the appointment of Philip Chun as our Technical Sales Representative for Toronto and Southwestern Ontario . Philip has a Bachelor of Science degree from the University of Waterloo as well as 9 years of experience in academic and industry research laboratories.
We are very excited to have Philip as part of our team.
If you would like to contact Philip, he can be reached via our toll free number at 1-888-593-5969 x 401 or by e-mail at philipc@biolynx.ca.
NuGEN and Hamilton Create Automated, Total target Prep Solution to Increase Throughput and Improve Gene Expression Analysis Workflow.
NuGEN
and the Hamilton Company, a world leader in precision liquid
handling announced the two companies have assembled an automated,
total target preparation solution to address the high-throughput
challenges required in processing large sets of clinical
samples for global gene expression analysis. The validated
solution, consisting of NuGEN's popular Ovation® Systems
and the Hamilton MICROLAB STARlet bench-top liquid handling
system, was carefully engineered to increase throughput
and improve workflow for small RNA samples commonly used
in clinical signature and biomarker discovery.
This fully integrated solution manages a full range of target preparation functions—RNA amplification, purification, fragmentation, and labeling—for gene expression analysis using Affymetrix GeneChip® and HT arrays. The net result of its improved workflow includes not only increased throughput, but higher reproducibility, reduced hands-on intervention, and the ability to process RNA from a wide spectrum of clinical samples including whole blood, biopsies, and laser-captured micro-dissected specimens.
