Many Thanks to Give

We would like to thank our customers for choosing Allele’s products and services. The economic conditions have been challenging for the last two years and are still difficult. Allele Biotech could not have possibly achieved what it has within this time period without the support of its customers.

We thank online readers who visit our Blogs, News and Technical Forum to make our webpage one of the highest ranked among biology reagent suppliers; surpassing Clontech, Stratagene, Promega, IDT, etc. (by online ranking service Alexa’s accounts).

We thank our collaborators (some converted from customers) and business partners (manufacturers, distributors, licensors) for working with Allele and making our plan of one new product per week a reality. Allele Biotech has a very innovative and able research team, but working with researchers outside of the company is always a major part of the R&D effort at Allele.

Allele Biotech is obliged to its employees for their creativity, organization, dedication, and professionalism. The culture that they have nurtured here is to be individually motivated as a basic research group, and to be disciplined and organized as a service provider at the same time.

Allele Biotech is a beneficiary of the US government’s policies on supporting biomedical research and innovation. We appreciate the support for our basic research in the forms of grants and contracts from the NIH and the IRS, and ultimately the American tax payers. It is our duty to create and produce better tools for improving human health.

Thank you once again and have a Happy Holiday!

New Product of the Week 112210-112810:

Photoconvertible FP mClavGR (green-to-red) on lentiviral vector pLICO, email FP@allelebiotech.com for product details.

Promotion of the Wee 112210-112810:

10% additional discount off HPLC siRNA’s promotion price, $135/15 nmol, use promo code FB112210si, order by emailing oligo@allelebiotech.com.

Tags: Clontech, collaboration, collaborators, customer opinions, customer service, IDT DNA, IDT oligo, Life Technologies, mClavGR, photoconvertible FP, promega, Stratagene, thanksgiving

Features of Allele’s Virus Packaging Services

High Titer
Allele Biotech’s HiTiter™ Viral Packaging Services, using proprietary technologies that are unique and highly efficient, can easily yield 10e8 to 10e9 TU/ml lentivirus/retrovirus without concentrating steps. With a concentration step 10e9 to 10e10 TU/ml can be achieved, which is ideal for in vivo research. It’s the world’s most powerful viral packaging platform.

Quick Turnaround
Allele Biotech has consolidated its procedure of HiTiter™ Viral Packaging Services as a standard production-line resulting in a quick turnaround. Typically, a standard 10e8 TU/ml 2ml packaging service is completed in one week and delivered in less than two weeks. You send us virus transfer plasmids, wait a week or two, and receive high titer ready-to-use virus. Imagine how our ingenuity and innovation can pave the way for your research!

Ideal for Complicated Constructs
Viral packaging, especially for complicated constructs, has been intrinsically difficult for most researchers. Now with Allele Biotech HiTiter™ Viral Packaging Services, you have an ideal option for packaging even the most demanding constructs! We have successfully developed the 4-In-1 iPSC Generation Lentivirus, expressing 4 genes (hOct3/4, hSox2, hKlf4 and hc-Myc) in one lentiviral construct, which has been very popular in the filed of stem cell research.

Fluorescent Proteins & Drug Resistance
Allele Biotech and our collaborators design, evolve, and select new FPs that suit different applications. These brighter fluorescent proteins, sometimes together with drug resistance genes, have been incorporated into our lentiviral/retroviral systems. When you order our viral packaging service, you will automatically receive a 20% discount for all our listed viral plasmids.

Full RNA Interference Services
We have RNAi lentiviral packaging services, RNAi validation services, and RNAi screening services, which can meet different demands in the filed of RNA Interference. The RNAi lentiviral packaging services include Gene-To-Silence™, Sequence-to-Virus™ and DNA-to-Virus™. The plasmids we use for these services have build-in FPs and drug resistance genes, convenient for virtually all research purposes.

New Product of the Week 111510-112110:

Cre/loxP Reporter Cell Line LoxP-lacZ Human Fibroblast, ABP-RP-CLACLOX.

Promotion of the Wee 111510-112110:

Fine agarose for DNA or RNA gel, buy 500g, get a second bottle of agarose at 40% off, use promo code FB111510AR, order by emailing oligo@allelebiotech.com

Tags: custom viral packaging, high titer, lenti-cre, lentivirus, packaging lentivirus, pre-packaged viruses, retrovirus, RNAi screening

Integration of Orbigen Products

Allele Biotech has been marketing products previously offered by Orbigen since July of 2008. In the process, Allele Biotech has transferred the majority of the Orbigen products to its own product lines, and left out a few that were no longer in demand, under proper license, or in some cases replaced with newer and better versions.

The biggest class of Orbigen products now under the Allele Biotech brand name are the polyclonal antibodies such as anti-EDG1 EDG2… EDG8 (catalogue numbers: PABs 10626, 10619, 10628, 10630, 10496, 1063), anti-BMRPs (catalogue numbers: PABs 10536, 10537, 10538). Other popular polyclonal antibodies from Orbigen include PAB-10983, PAB-10216, PAB-10683, PAB-11651, PAB-11141, PAB-10241, PAB-10469, PAB-10778, PAB-11248, PAB-11665, PAB-10563, PAB-10774. Orbigen antibodies have been tested in hundreds to thousands of labs over more than ten years.

One key product group developed by Orbigen is the baculovirus system, which includes the viral Sapphire genomic DNA and pOrb transfer vector plasmid for efficient packaging. We have further developed the pOrb vector into a bicistronic vector for dual expression or mammalian infections.

Retrovirus packaging cells, on the other hand, have been obtained from non-Orbigen sources and are now called Phoenix 2 as products under a new product line, the Gryphon retrovirus system. These cells have similar functions but were built by different researchers and further selected at Allele Biotech. Like all packaging cells we have experience with, these cells do not attach well after a freeze/thaw cycle. It is absolutely essential to wash away DMSO and better seed them in high quality, attachment enhancing dishes such as our EcoCulture plates.

Products previously under the Orbigen brand name are now included in Allele Biotech’s brand new shopcart (now beta-testing at shop.allelebiotech.com). We hope the new functionalities at the new site combined with the relevant knowledge and information you used to enjoy at our current allelebiotech.com site will provide you with an improved shopping experience.

New Product of the Week 110810-111410:

Retroviral vector with drug resistance new version, ABP-PVL-IRES10P.

Promotion of the Week 110810-111410:

GFP-Trap beads-immobilized camelid antibody for GFP fusion pull down. 15% off ACT-CM-GFA0050, any number of vials ordered this week. Use code SP110810.

Tags: Allele Biotech Orbigen merge, anti-BMRP, anti-EDG, biocarta antibodies, Orbigen, Orbigen antibodies, PAB-10, PAB-11, polyclonal antibodies

Cell Based Assays in Cell Cycle II

GFP-HDHB(CT)/RFP-HDHB(CT)
The C-terminal region of human DNA helicase B (HDHB) is a 99 amino acids polypeptide that contains both a putative nuclear localization signal (NLS) and a nuclear export signal (NES). It undergoes cell cycle-dependent translocation. GFP-HDHBCT is a fusion of GFP and the C-terminal region of HDHB. It remains inside the nucleus during G1 phase, then translocates into and remains in the cytoplasm during S and G2 phases. After nuclear envelope breaks down in M phase, this biosensor is localized throughout the cell. After nuclear envelope reformation at the end of M phase, the G1 phase biosensor is imported into the nucleus.

Hahn AT, Jones JT, Meyer T. Quantitative analysis of cell cycle phase durations and PC12 differentiation using fluorescent biosensors. Cell Cycle. 2009 Apr 1;8(7):1044-52. Epub 2009 Apr 2.

PM-YFP-NLS
This biosensor is a fusion protein comprising three components, a reversible plasma membrane–targeting domain fused to the N terminus of enhanced yellow fluorescent protein (EYFP), which is in turn fused to the N terminus of a nuclear localization signal (NLS). The nuclear localization signal would anchor the reporter’s localization to the nucleus during the G1, S and G2 phases (interphase) of the cell cycle. Upon the nuclear envelope breakdown at the onset of prometaphase, the plasma membrane–binding domain would cause reversible translocation of the biosensor to the plasma membrane where the fluorescence could be monitored by microscopy.

Jones JT, Myers JW, Ferrell JE, Meyer T. Probing the precision of the mitotic clock with a live-cell fluorescent biosensor. Nat Biotechnol. 2004 Mar;22(3):306-12.

E2F-mClaverGR TRE Reporter
The E2F family of transcription factors is a key regulator of cell-cycle checkpoints in mammalian cells. The E2F protein is a major target of the retinoblastoma gene product (Rb) and the activity of E2F/pRb is intimately connected with the G1-S transition of the cell cycle. The E2F protein forms a heterodimer complex with DP1, which binds to E2F response elements and initiate transcription.
The E2F-responsive mClaverGR construct encodes a photoconvertible green-to-red fluorescent protein reporter gene under the control of a minimal (m)CMV promoter and tandem repeats of the E2F transcriptional response element (TRE). We have experimentally optimized the number of response elements as well as the intervening sequence between response elements to maximize the signal to noise ratio.

Ki67p-GFP
Ki67, encoded by MKI67 gene, is expressed in all phases of the active cell cycle (G1, S, G2 and M phase), while is absent in the resting phase (G0). Therefore, it is routinely used as a marker of cell cycling and proliferation. Recently, Zambon AC[1] cloned and characterized the 1.5 kb proximal promoter (Ki67p) of the human Ki67 gene. A reporter, Ki67p-GFP, was further constructed to express GFP under Ki67p. Their data verified that GFP driven by Ki67p is co-expressed in cells with endogenous Ki67 and is correlated with cells transitioning through G1/S/G2/M phases of the cell cycle. Mitomycin C induced G1/S/G2/M blocking or cell-density induced cell cycle arrest both attenuate Ki67p activity.

Zambon AC. Use of the Ki67 promoter to label cell cycle entry in living cells.Cytometry A. 2010 Jun;77(6):564-70.

New Product of the Week 110110-110710:

BacMam virus for subcellular localization marker-mWasabi fusion constructs. Email FP@allelebiotech.com for details.

Promotion of the Week 110110-110710:

Oct3/4-promoter-RFP pluripotency reporter lentivirus. 20% off ABP-SC-ROCT4G1$149.00 using code SP110110.

Tags: cell assays, cell cycle, G1 phase G2 phase, interphase, M phase, nuclear envelop, nuclear localization, S Phase