October 2015 News We add two antibodies to the Heat Shock 60 (HSP60) protein, a major component of the mitochondrial membrane. These are the mouse monoclonal MCA-1C7 and a rabbit polyclonal RPCA-HSP60. Both are very clean and specific on western blots of crude extracts of cells and tissues and both stain mitochondria beautifully, as shown here. We release a new chicken antibody to 2′,3′-cyclic nucleotide 3′-phosphodiesterase (CNP), CPCA-CNP, an excellent marker of oligodendrocyes and their processes. This complements our previously released rabbit and mouse antibodies to this protein, MCA-3H5 and RPCA-CNP. We release an antibody binding to the C-terminal netrin-like domain of complement C3, MCA-5F2, the first of a series of antibodies to different regions of this large and important serum protein. This antibody can be used to track the α subunit of C3 and its many proteolytic fragments on western blots. We also release a monoclonal antibody to phospho-MECP2, MCA-2E2, which recognizes this important transcriptional regulator if it is phosphorylated on Ser421. Finally, we attended the Society for Neuroscience 2015 meeting in Chicago, where we displayed our wares in booth 1870, easy to remember as that is also the date of the Franco-Prussian war. We showed some of our immunocytochemical and immunohistochemical images and were pleasantly surprised at how popular they were, we sold well over one hundred. Some of these were high quality 24″ X 36″ glossy prints which are available for $25 and can be ordered online. We also displayed numerous 20″ X 29″ prints of the same images which we had produced by a wrapping paper company. These were very cheap to print so we only asked $5 for them- however they look great when framed. Since these are so cheap shipping would be a significant part of the total cost for online orders. As a result we have made a package of the 8 most popular of these, Poster-Col-1, which can be purchased for $40, shipping included.
September 2015 News We release a novel mouse monoclonal antibody specific for microtubule associated protein 2D (MAP2D), MCA-2C4. This was raised against the purified recombinant human MAP2D, an embryonic form of this major CNS molecule. Our experiments show that this antibody also binds to MAP2C, the smallest product from the MAP2 gene. The antibody therefore will stain all forms of MAP2, not just the high molecular weight mature forms which are recognized by antibodies to the MAP2 “projection domain”, such as our MCA-4H5 and MCA-5H11. The antibody also works very well on aldehyde fixed sections of brain tissue. We also add a bunch of new pure recombinant proteins to our growing collection. The first is a purified recombinant form of a green fluorescent protein (GFP), catalog Prot-r-aceGFP. GFP was originally isolated from a jellyfish and it and its variants have become widely used as a tracer in many experimental paradigms. We also add a chicken polyclonal antibody raised against this GFP preparation, CPCA-GFP, which can be used to verify the size of GFP fusion proteins on western blots and to amplify the GFP signal in transgenic, tranfected and transduced tissues and cells. Another fluorescent protein is the intriguing EosFP protein, Prot-r-EosFP, another green fluorescent protein distantly related to GFP but originally isolated from a coral. This has the interesting property of changing to a red fluorescent protein following irradiation with blue or UV light. This property makes it very useful for pulse chase type experiments in live cells. Another GFP, this time from a different coral is FP506, Prot-r-FP506. These three proteins complement our recombinant mCherry, Prot-r-mCherry preparation. Antibodies to all of these proteins are either in preparation or already available. Our products are now listed on the CiteAb site so you can rapidly find peer reviewed publications which make use of our products. This only works if the antibody was purchased from us directly, many of our antibodies have very large numbers of citations but were purchased through our many OEM partners and so appear on their CiteAb pages and not ours. Also check out this JNNP article “Are neurofilaments heading to the ALS clinic?”, discussing the potential utillity of the measurement of blood and CSF levels of neurofilament proteins as diagnostics and outcome measures for ALS. This is an approach we pioneered in the EnCor lab and with our clinical and academic colleagues. We also provide a link to our founders blog, which will contain his rantings and ravings about a variety of topics, some scientific, some not. This can be accessed at GerryShawBlog. If you really want you can also get an RSS feed on this stuff here.
August 2015 News We release three new antibodies to 2′-3′-cyclic nucleotide phosphodiesterase (CNP), a major protein of oligodendrocytes in the central nervous system. Such antibodies are therefore excellent markers of oligodendrocytes, and, in the peripheral nervous system, Schwann cells. There are two mouse monoclonals MCA-1H10 and MCA-3H5 and a rabbit polyclonal, RPCA-CNP. All work strongly and specifically on blots and all can be used to identify oligodendrocytes in culture and in sections. We also release a new mouse monoclonal antibody to parvalbumin, a small calcium binding protein found in muscle and is certain neurons in the central nervous system. This antibody is MCA-3C9, and we also show that is does not cross-react with the related proteins calretinin or calbindin. We also release a novel mouse monoclonal antibody, MCA-6C6, to calreticulin, an important protein implicated in both calcium buffering and autophagy. This antibody works well on cells in tissue culture and on western blots and epitope for the antibody is the LIR peptide, which the the binding site for LC3 and Atg8 family proteins. Finally, on the 28th of August we presented some of our cell and tissue images at the Gainesville Arts and Crafts Show for the first time. This is monthly event and here is the Facebook page the show made about us. Although it was windy and rained, as it does this time of year in North Central Florida, this went surprisingly well, apparently pictures of antibody stained brain cells may have some artistic and/or commercial merit.
July 2015 News We add yet more antibodies and recombinant constructs to our catalog. We release a rabbit polyclonal antibody to total MECP2, RPCA-Mecp2. MECP2 is an important transcription factor with particular activity in the nervous system. We have phosphospecific antibodies of MECP2 under development. We release an rabbit antibody to Green Fluorescent Protein (RPCA-GFP), which can be used to enhance the GFP signal or verify that a construct of the appropriate size has been expressed. We release a new chicken antibody to parvalbumin, CPCA-Pvalb. Parvalbumin is a small calcium binding protein concentrated in muscle and in certain CNS neurons. We also release a collection of recombinant human MAP2 protein constructs, covering almost all regions of this large and important molecule. These constructs are Prot-r-MAP2C, Prot-r-MAP2D, Prot-r-MAP2-P1, Prot-r-MAP2-P2 and Prot-r-MAP2-P3. The constructs were prepared using the human DNA sequence which was codon optimized for expression in E. coli, which was used to prepare bulk amounts of these proteins. These are suitable for use as ELISA standards, for epitope mapping of antibodies or for the generation of novel region specific antibodies.
May 2015 News We release more new antibodies. To complement our calbindin antibodies we now release a new mouse monoclonal antibody to calretinin. Both calbindin and calretinin are small calcium binding proteins which are each found in subsets of neurons in the CNS, and antibodies to them are therefore useful for the identification of specific neuronal cell types. Our new mouse monoclonal antibody to calretinin is MCA-3G9. We also introduce an antibody to fibrillarin made in chicken, CPCA-Fib, an excellent marker of nucleoli in a wide variety of cell types, This is an excellent complement to our widely used fibrillarin/Nop1p monoclonal MCA-38F3.
More February News: Our immunocytochemical images continue to be used in books, journals and news articles, usually without any attribution, since we did not bother to copyright or watermark any of them. Many have been sent freely to our OEM partners and in some cases to Wikipedia. Scroll down in this very recent high profile on line article entitled “An Inflammatory Theory of Brain Disease” to see one of our images. The article is an interesting PBS NOVA write up of some progress (and also lack thereof) in the search for effective therapies to treat human brain diseases. The image in the article is of our mixed neural cells grown in tissue culture stained with our microglial cell marker Coronin 1a in green and the neuronal marker α-internexin in red. This particular image was in fact provided to Wikipedia Commons for unfettered use as shown here.
February: We release several new antibodies. These are a new mouse antibody to muscleblind like protein 1, MCA-1H1, and several to Calbindin, made in both mouse MCA-5A9, MCA-4H7 and chicken CPCA-Calb (page in preparation). We also continue to develop a better understanding of all of our current reagents. For example, we previously made antibodies to the abundant cytoplasmic enzyme Aldolase C, both monoclonal and polyclonal. A potential problem with antibodies to this protein is cross-reactivity with the Aldolase A and B molecules which are very similar in amino acid sequence. Each Aldolase is 70-80% identical to the other two and the molecules have very similar molecular sizes on SDS-PAGE gels so one cannot distinguish which protein is being detected using western blotting. So we went to the trouble of expressing both human Aldolase A and B in order to test our panel of Aldolase C antibodies. We found that two of our monoclonals MCA-1A1 and MCA-4A9 were absolutely specific for Aldolase C, while a third MCA-E9 recognized all three Aldolases. We suspected that this might be the case since MCA-1A1 and MCA-4A9 were made against the C and N-terminal peptides respectively, which are the least conserved parts of the three Aldolases. In contrast, MCA-E9 was made against the intact human recombinant protein, and we now know that the epitope does not include either the N or C terminal sequences. Our rabbit polyclonal antibody to Aldolase C, RPCA-AldC, (page in preparation) not surprisingly, also recognizes all three Aldolase enzymes. Since Aldolases are abundant enzymes found in many tissues, we believe that using antibodies such as MCA-E9 and RPCA-AldC, which recognize all three, might be particularly useful as a western blotting standard, similar to our pan-specific actin antibody MCA-5J11.
January: We went to the Society for Neuroscience 2014 meeting in Washington DC a few weeks ago, and presented our new antibodies, ELISAs, recombinant proteins and other products, which generated some considerable interest. However the most interest seemed to be directed at our collection of high resolution images which we presented as 24″X36″ posters. We sold some of these and we had mentioned that we could also print these for use as Christmas wrapping paper. Well, we did this and the paper looks very nice, almost as nice as the posters. So we are now offering these for sale at very reasonable prices. One sheet of 20″ X 29″ paper is $3.50, five is $14.00 and ten is $25.00. Shipping will be $6.00, irrespective of the number you order. We currently have two of these available, identical to our posters Image-2 and Image-7. We realize that it is too late for Christmas this year, but there is always next year, birthdays etc. The images are of high enough quality, despite the price, to make nice posters or wall decorations. We don’t have these two items in our on-line shopping cart yet, so just email us at admin@encorbio.com if you are interested in these.
