Social Media for the Consumer

Business has become an ever changing, dynamic landscape where the power and information no longer lies strictly in the hands of “the right people.” In less than a decade Facebook has managed to link nearly a billion people from tech savvy teens to curious senior citizens. Similarly, Twitter sees over 340 million tweets that reaches over 500 million active users per day. Social media is a wide reaching technology with infinite possibilities that has been synthesized and is fueled by people, not corporations.

In a recent Industry report, 93% of businesses reported using social media as a marketing tool with the benefits being increased exposure and customer interaction (Stelzner, 2011). What does this study mean to consumers? Businesses are coming to you and responding to your feedback. Social media gives you, the consumer; back the power and the opportunity to benefit from your loyalty. Most businesses, from restaurants to labs, offer weekly tips or discounts on their services to the customers who “follow” or “like” them. Here at Allele Biotech, we have begun offering special weekly promotions, information about our products and interesting biotech news to our customers who either “follow” us on Twitter or “like” us on Facebook. For the entire month of August we are offering 15% off just for a “like” or “follow.” If you are one of the billion users, it doesn’t take much more than a click for instant rewards.

We recognize that the reason we create this technology and improve is because of the consumers. Allele Biotech would like to thank you for your loyalty and encourage you to remain active within our social media to push us forward and to help us continuously strive for more. With the creation and exponential popularity of social media, the power and information as become available to the most important people, the consumers.

Twitter @Allele_Biotech
Facebook http://www.facebook.com/pages/Allele-Biotechnology-and-Pharmaceuticals-Inc/78331924957

Picture Blog — Making mRNAs by In Vitro Transcription for Transgene Expression and R-iPSCs

R-iPS Cell FAQ 2:
What is the expected yield from the in vitro trancription (IVT) reactions?

Performed as described, you should recover around 40 ug RNA from each 40 uL IVT reaction.

R-iPS Cell FAQ 3:
How can the success of the RNA synthesis protocol be assessed?

Run 500 ng (5 uL) of the concentration-adjusted products on an E-gel to check for consistent product yield and relative product sizes, and to confirm the absence of secondary bands or smears.

Tags: feeder-free, footprint free, huma iPS, IVT, mRNA, R-iPSCs, transgene, Warren

Record number of papers citing the GFP-Trap group products in 2011

The following are references in regards to GFP Trap published in the second half of 2011 (not a complete list); a high quality GFP-binding protein based on a single domain antibody derived from Camelids. It is characterized by a small barrel shaped structure (13 KDa, 2.5nm X 4.5 nm) and a very high stability (stable up to 70°C, functional within 2M NaCl or 0.5% SDS). With much greater stability, specificity, and affnity, GFP-Trap®, the recent addition to antibodies for immunoprecipitation, should make GFP the most suitable tag for immunoprecipitation assays.

For live PubMed links, view this version please.

Krastev, D. B., Slabicki, M., et al. (2011). A systematic RNAi synthetic interaction screen reveals a link between p53 and snoRNP assembly. Nature Cell Biology. 13: 809-818. PubMed

Aboobakar, E. F., Wang, X., et al. (2011). The C2 domain protein Cts1 functions in the calcineurin signaling circuit during high temperature stress responses in Cryptococcus neoformans. Eukaryotic Cell. EC. 05148-05111v05141. PubMed

Uhrig, R. G. and Moorhead, G. B. G. (2011). Two ancient bacterial-like PPP family phosphatases from Arabidopsis thaliana are highly conserved plant proteins that possess unique properties. Plant Physiology. PubMed

Larance, M., Kirkwood, K. J., et al. (2011). Characterization of MRFAP1 Turnover and Interactions Downstream of the NEDD8 Pathway. Molecular & Cellular Proteomics. PubMed

Hattersley, N., Shen, L., et al. (2011). The SUMO protease SENP6 is a direct regulator of PML nuclear bodies. Molecular Biology of the Cell. 22: 78-90. PubMed

Rancz, E. A., Franks, K. M., et al. (2011). Transfection via whole-cell recording in vivo: bridging single-cell physiology, genetics and connectomics. Nature Neuroscience. 14: 527-532. PubMed

Palmer, C. S., Osellame, L. D., et al. (2011). MiD49 and MiD51, new components of the mitochondrial fission machinery. EMBO reports. 12: 565-573. PubMed

Pichler, G., Wolf, P., et al. (2011). Cooperative DNA and histone binding by Uhrf2 links the two major repressive epigenetic pathways. Journal of Cellular Biochemistry. 112: 2585-2593. PubMed

Mitchell, L., Lau, A., et al. (2011). Regulation of Septin Dynamics by the Saccharomyces cerevisiae Lysine Acetyltransferase NuA4. PLoS One. 6: e25336. PubMed

Engeland, C. E., Oberwinkler, H., et al. (2011). The cellular protein Lyric interacts with HIV-1 Gag. Journal of virology. JVI. 00174-00111v00171. PubMed

Wang, C. and Youle, R. (2011). Predominant requirement of Bax for apoptosis in HCT116 cells is determined by Mcl-1’s inhibitory effect on Bak. Oncogene. PubMed

Tulloch, L. B., Howie, J., et al. (2011). The inhibitory effect of phospholemman on the sodium pump requires its palmitoylation. Journal of Biological Chemistry. 286: 36020-36031. PubMed

Sun, L. and Wang, C. C. (2011). The Structural Basis of Localizing Polo-Like Kinase to the Flagellum Attachment Zone in Trypanosoma brucei. PLoS One. 6: e27303. PubMed

Bouttier, M., Saumet, A., et al. (2011). Retroviral GAG proteins recruit AGO2 on viral RNAs without affecting RNA accumulation and translation. Nucleic acids research. PubMed

Matos, J., Blanco, M. G., et al. (2011). Regulatory Control of the Resolution of DNA Recombination Intermediates during Meiosis and Mitosis. Cell. 147: 158-172. PubMed

Nagel, C. H., Albrecht, N., et al. (2011). Herpes Simplex Virus Immediate-Early Protein ICP0 Is Targeted by SIAH-1 for Proteasomal Degradation. Journal of virology. 85: 7644. PubMed

Studencka, M., Konzer, A., et al. (2011). Novel roles of C. elegans heterochromatin protein HP1 and linker histone in the regulation of innate immune gene expression. Molecular and Cellular Biology.PubMed

Muehlen, S., Ruchaud-Sparagano, M. H., et al. (2011). Proteasome-independent Degradation of Canonical NFŒ?B Complex Components by the NleC Protein of Pathogenic Escherichia coli. Journal of Biological Chemistry. 286: 5100. PubMed

Galan, J. A., Paris, L. L., et al. (2011). Proteomic Studies of Syk-Interacting Proteins Using a Novel Amine-Specific Isotope Tag and GFP Nanotrap. Journal of the American Society for Mass Spectrometry. 1-10. PubMed

Chamousset, D., De Wever, V., et al. (2010). RRP1B Targets PP1 to Mammalian Cell Nucleoli and is Associated with Pre-60S Ribosomal Subunits. Mol Biol Cell. PubMed

Kovacs, E. M., Verma, S., et al. (2011). N-WASP regulates the epithelial junctional actin cytoskeleton through a non-canonical post-nucleation pathway. Nature Cell Biology. 13: 934-943. PubMed

Boysen, K. E. and Matuschewski, K. (2011). Arrested oocyst maturation in Plasmodium parasites lacking type II NADH: ubiquinone dehydrogenase. Journal of Biological Chemistry. 286: 32661-32671. PubMed

Mortusewicz, O., Fouquerel, E., et al. (2011). PARG is recruited to DNA damage sites through poly (ADP-ribose)-and PCNA-dependent mechanisms. Nucleic acids research. 39: 5045. PubMed

Graewe, S., Rankin, K. E., et al. (2011). Hostile takeover by Plasmodium: reorganization of parasite and host cell membranes during liver stage egress. PLoS Pathogens. 7: e1002224. PubMed

Yang, X. D., Huang, S., et al. (2011). Distinct and mutually inhibitory binding by two divergent Œ?-catenins coordinates TCF levels and activity in C. elegans. Development. 138: 4255-4265. PubMed

Pollithy, A., Romer, T., et al. (2011). Magnetosome expression of functional camelid antibody fragments (nanobodies) in Magnetospirillum gryphiswaldense. Applied and environmental microbiology. 77: 6165-6171. PubMed

Kozubowski, L., Thompson, J. W., et al. (2011). Association of Calcineurin with the COPI Protein Sec28 and the COPII Protein Sec13 Revealed by Quantitative Proteomics. PLoS One. 6: e25280. PubMed

Garcia-Gomez, J. J., Lebaron, S., et al. (2011). Dynamics of the putative RNA helicase Spb4 during ribosome assembly in Saccharomyces cerevisiae. Molecular and Cellular Biology. 31: 4156-4164. PubMed

Van Damme, D., Gadeyne, A., et al. (2011). Adaptin-like protein TPLATE and clathrin recruitment during plant somatic cytokinesis occurs via two distinct pathways. Proceedings of the National Academy of Sciences. 108: 615. PubMed

Qvist, P., Huertas, P., et al. (2011). CtIP Mutations Cause Seckel and Jawad Syndromes. PLoS Genetics. 7: e1002310. PubMed

Labella, S., Woglar, A., et al. (2011). Polo Kinases Establish Links between Meiotic Chromosomes and Cytoskeletal Forces Essential for Homolog Pairing. Developmental Cell. PubMed

Harterink, M., Port, F., et al. (2011). A SNX3-dependent retromer pathway mediates retrograde transport of the Wnt sorting receptor Wntless and is required for Wnt secretion. Nature Cell Biology. 13: 914-923. PubMed

Konopacki, F. A., Jaafari, N., et al. (2011). Agonist-induced PKC phosphorylation regulates GluK2 SUMOylation and kainate receptor endocytosis. Proceedings of the National Academy of Sciences.PubMed

Chuhma, N., Tanaka, K. F., et al. (2011). Functional connectome of the striatal medium spiny neuron. The Journal of Neuroscience. 31: 1183-1192. PubMed

Jackson, B. R., Boyne, J. R., et al. (2011). An Interaction between KSHV ORF57 and UIF Provides mRNA-Adaptor Redundancy in Herpesvirus Intronless mRNA Export. PLoS Pathogens. 7: e1002138. PubMed

Tags: anti-GFP, camelid antibodies, Chromotek, Co-IP, co-IP 101, GFP-Trap, GFPTrap, immunoprecipitation, nAb: Camelid Antibodies, nanobodies, pulldown, RFPTrap, VHH, VHH antibody

Year-end message from Allele Biotech

The year 2011 has been an exciting and eventful year for many people. Throughout the year, we have been working diligently to bring the best research methods in many areas to our fellow researchers through innovation and entrepreneurship. Thanks in part to the government’s stimulus and grant support in 2011, we established several new product lines, including the Stealth iPS induction mRNA templates and reagents, a great new photoconvertible fluorescent protein in mClavGR2 (through collaboration with academic colleagues), and a highly efficient lentivirus-based shRNA packaging service as a result of an NCI SBIR contract.

As you all must have noticed by now, in July we redesigned our website to present our products in an easier, more user friendly manner, while adding a convenient online purchasing system. We have received a lot of positive feedback from customers telling us how “cool” the new site is, and how easy it is to use and redeem promotions. Towards the end of the year, our dedicated marketing and sales teams reinstated our biweekly email newsletters (to receive our messages on new discoveries and technologies, or be the first to use our promotions, sign up online under “Newsletter”).

All of these efforts would have been meaningless without our customers, who ultimately gave us the opportunity to be in the business we love and are trained to do. By selecting our products, sending us feedback, and “retweeting” or “reposting” our messages, you have been tremendously valuable to every one of us here at Allele. We thank you from the bottom of our hearts. In return, we will continue to invest and do our very best to provide new tools for advancing your research. Watch for our brand new monomeric fluorescent protein that can be nearly 10 times brighter than EGFP; a more powerful iPSC generation method that could potentially reprogram in just a few days, and much much more in 2012!

Tags: 2012, EGFP, iPS, iPSC, mRNA templates, NCI SBIR, new fluorescent proteins, reposting, retweeting

Making Recombinant Glycosylated Proteins (I)

Many mammalian cell membrane-bound and secreted proteins are glycosylated. The degree of their modifications may be dependent upon tissue specificity and cellular states such as normal vases cancerous. The existence of these proteins in bodily fluids, in combination to their relevance to diseases, makes glycosylated proteins good candidates for clinical diagnostics. Understanding the biogenesis, structure, and functions of these proteins will also aid research and prevention of cancers.

Cancer formation is a heterogeneous and complex process, involving many factors and cellular signaling pathways in each type of cancer. There are more than 1,200 potential cancer biomarkers identified in the literature by a 2006 review. We found that ~ 70% of the 1,261 proteins listed in are naturally secreted proteins, and some 40-50% glycosylated. The addition of carbohydrate groups during protein glycosylation to asparagines (N-linked), or threonines or serines (O-linked) residues may result in mono-, disaccharide- or branched oligosaccharide composed of as many as 20 monosaccharide residues. Glycosylation, together with other modifications, often change the apparent molecular mass of a secreted protein to many folds to that predicted by amino acid sequence. Such heavy modifications on the surface of proteins can influence their functions as well as characteristics as antigens or analytes. Studies of glycosylated proteins offer great opportunities for improving cancer diagnostics.

There are increasing demands for these glycosylated human proteins in good quantity, purity and affordability by the scientific community to perform fundamental and clinical studies in relation to cancer. Such proteins cannot be expressed in bacteria or yeast because those cells do not carry out equivalent post-translation modifications (PTM) as in mammalian cells. Although there have been successful attempts to modify yeast cells to produce proteins with certain types of glycans attached, they were designed for expressing a few pharmaceutical proteins and not suitable for expressing a wide variety of cancer markers. Aside from PTM, expressing human proteins in microorganisms may be hindered by their different codon usage preferences and protein folding tendencies.

Surface Bind Just-a-Plate™ 96 PCR Purification kit, ABP-PP-JA100S, $78.

10% off Gryphon Packaging Cell Line for Retrovirus. Email oligo@allelebiotech.com along with promo code GP012411 when shopping at shop.allelebiotech.com.

Tags: cancer markers, glycosylated proteins, glycosylation, recombinant proteins, serum samples