Lentivirus-lanYFP Give Away

Promotion of the week 03-22-10 to 03-28-10: To help researchers get familiar with pre-packaged lentivirus, we offer free high titer lentivirus carrying a truly bright and fast maturing lanYFP (lancelet FP, new exclusively from Allele).  Infect virtually any mammalian cells by a single manipulation (pipeting) and watch cells turn green/yellow in about a day under microscope or on FACS.  Primarily a yellow FP, lanYFP will show brighter fluorescence than EGFP even when observed using standard GFP/FITC filter set.

New product of the week 03-22-10 to 03-28-10: anti-mTFP1/mWasabi polyclonal antibodies.  It is tailored-made for Alleleustrious mTFP1 and mWasabi, the brightest teal and green FPs.

Tags: EGFP, Fluorescent proteins, GFP, Lancelet, lentivirus, mTFP1, mWasabi, pre-validated virus, YFP

Fluorescent Protein-Based Assay Development II

FPs as pH and redox sensors:

The uses of FPs extend well beyond simple expression and fusion reporters.  While pH sensitivity (usually quenching of fluorescence by acidic pH) is generally considered a drawback for fusion tagging, it becomes a useful property for constructing pH sensors.  FPs specifically engineered to take advantage of pH sensitivity (“pHluorins”) report pH as either a change in fluorescent intensity or a change in the ratio of excitation at two different wavelengths, and may be used to monitor processes such as endocytosis or other pH-variable processes.  In such an application, the pH-sensitive FP is fused to a localization tag for the compartment of interest which experiences variable pH.  This technique can be used, for example, to visualize release of neurotransmitter-containing vesicles.  In addition to pH-sensitive FPs, redox-sensitive Aequorea GFP variants have been produced (roGFPs and others) which produce similar changes in fluorescence intensity or excitation ratio when exposed to differing redox conditions or reactive oxygen species.

Sensors based on circularly permuted FPs:

Because FPs have such a compact and stable beta-barrel fold with N and C termini close together, it is possible to engineer circularly permuted variants which retain their fluorescent properties.  Studies on circular permutation of FPs have led to the development of several different sensors which take advantage of domains inserted into sensitive areas of the fluorescent protein backbone.  The most famous of these are the GCaMP calcium sensors, in which a calmodulin domain has been inserted into a loop in GFP, yielding a sensor that reports calcium concentration as a change in fluorescent intensity.  Other circularly permuted FP variants, such as cpVenus (a yellow Aequorea GFP variant), have found usefulness in improving FRET sensor dynamic range (see next section).

FRET sensors:

Fluorescence resonance energy transfer (FRET) is a quantum mechanical process that allows the transfer of excited state energy between two fluorophores when they are in close physical proximity.  Because this process operates with a strong distance (1/r^6) and orientation dependence (strongest when chromophore dipoles are parallel or antiparallel), it lends itself to the construction of highly sensitive reporters of biochemical activity.  In FP FRET, excited state energy from a higher-energy (shorter wavelength) “donor” fluorescent protein is transferred to a lower-energy (longer wavelength) “acceptor” FP, leading to sensitized fluorescent emission from the acceptor and reduced emission (quenching) from the donor.  By linking donor and acceptor FPs with a domain which changes conformation in response to a biochemical activity of interest, this activity is reported as a change in the ratio of sensitized emission to direct-excitation emission of the acceptor (or a simple ratio of donor and acceptor emission).  FRET sensors have been engineered to specifically sense a wide variety of activities, including many protein kinases, as well as small molecules such as Ca2+ and neurotransmitters.  While design of a new FRET sensor generally requires a great deal of optimization and trail-and-error, this class of probe is among the most powerful tools currently available for investigating live-cell biochemistry.

New Product of the Week 03-15-10 to 03-21-10: Oct4-Sox2 2-in-1 lentivirus ABP-SC-LVI2in1 for effective iPS generation link: http://www.allelebiotech.com/shopcart/index.php?c=132&sc=122.
Promotion of the Week 03-15-10 to 03-21-10: 5% off plate oligos at all scales! www.allelebiotech.com/allele3/Oligo_96Plate.php We are doing our “window promotion” again, during a hour-long window, get any Allele’s High efficiency competent cells at 30% regular price, the time will be announced tomorrow on our Facebook page.

Tags: animal models, biosensors, calmodulin, cell based assays, circular permutatioon, Fluorescent proteins, FP sensor, FRET, GCaMP, pH sensors, redox sensor

Submit Your Pictures and Enter to Win $100 in Cash

Allele Biotech is holding another fun contest to give back to our loyal supporters and everyone has the chance to win $100 in cash! We are looking for interesting and relevant pictures, illustrations, and computer generated art that pertains to any Allele Biotech Product or service to replace the flashes on our homepage. If it is chosen to be used we will give you $100 cash!

Check out our technical data sheets, read our past blogs, and even use your own lab experience and impressions with Allele Biotech Products to come up with your entry. There are over 1000 products to choose from so everyone should have lots of inspiration!

All you have to do to enter is submit your picture to our facebook inbox! The picture has to be of your own creation and/or one to which you own exclusive rights. Response time and prizes will take about a week.

Let your creative juices flow and you could get $100 in return. Your picture could be funny, serious, or even super nerdy!

Used to promote our Sapphire(TM) Baculovirus Expression Service

All photos, illustrations, and computer generated graphics (aka. “the picture”) submitted must be lawfully owned by entrants who submitted them. Pictures that are not chosen will not be used in any way by Allele Biotech. Winners may collect their prize via cash, check, or a one-time Allele credit of $100 good toward any Allele Biotech purchase. Allele Biotech has full rights to pictures selected for use in Allele advertising and winners must forfeit any future rights to the picture. Contest to run indefinitely and to be terminated at any time.

Tags: cash prize, contest, submit pics for cash, win $100

New Product for 1-18-10 to 1-24-10 High Throughput DNA Oligos!

Need a plate of high quality oligos fast and for a great price? Allele’s newest product can help…

Introducing High Throughput DNA Oligos!

Anywhere from 48 – 96 desalt oligos per plate

Oligos 20 – 50 bases long

Available in two synthesis scales:

25 nmol scale only 13 cents per base

50 nmol scale only 17 cents per base

Every oligo is strictly controlled for quality:

Oligo quality is verified using MALDI-TOF mass spectrometry or Electrospray Ionization Mass Spectrometry.

Shipped or hand delivered in 5 days!

Oligos can be provided normalized at a specific concentration with fixed or variable volumes. Concentration options depend on the oligo length, purification, and scale of synthesis being ordered.

No minimum number of plates per order required!

To order, simply email your oligo sequences and names, indicate 25 nmol or 50 nmol, concentration preference in excel or notepad format to oligo@allelebiotech.com A template will be posted online here soon, please check back.

Allele…Continuing to introduce cost efficiency to research!

Tags: DNA oligo, high throughput oligos, plate oligos, price for oligos, save on reasearch

10 Years of Allele Biotech

Facts about Allele’s 10 years in business:

Products

New product lines added in 2009: iPS cells, Camelid Antibodies, DNA synthesis chemicals, Recombinant Proteins

Highlights: HiTiter Lentiviral Systems, Baculovirus for Mammalian Expression (BacMam), Feeder Cells, shRNA on Viral Vectors, shRNA Validation FP Vector, ProperFold Protein Folding Vector, Validated AllHPLC synthetic siRNA

New Service Groups in 2009: Viral Packaging, RNAi Validation/Screening, FP-based Assay Development

Numbers

Since April, we have added at least one new product every week! We currently run one new promotion per week as well.

A bit of history–did you know that…

Allele Biotech obtained 5 NIH grants in its first three years since establishment. As a matter of fact, Allele Biotech was funded entirely by NIH grants

Allele filed its first patent application in its second year of operation, which was on DNA-driven RNAi and resulted in an outlicensing deal with Promega. As result of the applications, Allele has received 3 US patents on DNA-encoded shRNA, siRNA using promoters such as U6 and H1.

During the past 10 years, Allele was the first to sell U6-based RNAi vectors, the only supplier of bFGF-expressing feeder cells for iPSC, most likely a top 3 provider of baculovirus expression systems, camelid antibody products, iPS creating viral particles, and the most active commercial developer of fluorescent proteins.

Tags: 10 years in business, Allele Anniversary, camelid antibodies, NIH grants, oligos, RNAi