March 2018 News

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We finally get done with updating our web site, after several months of playing around with it, so we hope you like it!

We have made an excellent rabbit polyclonal antibody to IBA1 (a.k.a. AIF1), RPCA-IBA1. The IBA1 protein is heavily expressed in microglia but not in neurons, astrocytes, oligodendrocytes or other kinds of CNS cell. As a result, antibodies to IBA1 are widely used to identify and study microglia in CNS cultures and sections. Microglia are the immune cells of the CNS and have important roles in the cellular responses to a variety of CNS damage and disease states. We also have an excellent high resolution poster showing staining of microglial cells with this antibody on rat cerebellum, counterstained withe the neuronal marker MAP2, see poster-27. Here is an image of that.

We continue to characterize our antibodies in ever greater detail; for example we recently epitope mapped our two monoclonal antibodies to the intermediate filament protein vimentin, MCA-2A52 and MCA-2D1. This allows us to confidently predict which species these antibodies will work on. For another example see our new findings made on the widely used mouse monoclonal antibody to neurofilament NF-L MCA-DA2.

August 2016 News

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We release new mouse monoclonal antibodies to MECP2, the methyl CpG binding protein 2. This is an important transcriptional regulator of neurons sequence defects in which are associated with Rett syndrome, a serious neurodevelopmental disorder. These are the IgG2b MCA-5H12 and the IgG1 MCA-4E11. We release an excellent chicken antibody to the neural stem cell marker Nestin, CPCA-Nes. We also release a rabbit polyclonal antibody to fibrillarin, RPCA-Fib an excellent marker of nucleoli.

June 2016 News

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We release two more Cas9/CRISPR related antibodies, a rabbit polyclonal to Streptococcus pyogenes Cas9, RPCA-Cas9-AP and a mouse monoclonal, MCA-3F9 to the same protein. These can be used to confirm the expression of the most commonly used Cas9 molecule in transfection experiments.

April 2016 News

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We continue to release antibodies to Cas9, the key enzyme in the CRISPR gene editing system. These are RPCA-Cas9-SA and CPCA-Cas9-SA. These polyclonal antibodies recognize the Cas9 protein from Staphylococcus aureus, which is considerably smaller than the Streptococcus pyogenes derived Cas9. Since the two Cas9 enzymes are quite different in amino acid sequence our antibody to S. aureus does not bind S. pyogenes Cas9, and antibodies to the S. pyogenes form would not be expected to bind S. pyogenes Cas9. We also release a rabbit polyclonal antibody to the human neural stem cell marker Nestin, RPCA-Nestin. We release several biochemical preparations of major CNS proteins.  One is purified pig GFAP, Prot-m-GFAP-por, biochemically isolated from pig spinal cord. This preparation has post-translational modifications absent from our recombinant forms of the same protein based on the human and rodent sequence. These are our Prot-r-GFAP and Prot-r-GFAP-rat products respectively. We also release pure protein preparations of both bovine and porcine derived pNF-H, the phosphorylated axonal form of neurofilament NF-H. These are Prot-m-pNF-H-bov and Prot-m-pNF-H-por.

March 2016 News

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The life sciences have recently been revolutionized by the exploitation of the bacterial CRISPR system. CRISPR is an acronym for “Clustered Regularly Interspaced Short Palindromic Repeats”. The short palindromes are DNA sequences generated by the bacterial host, while the DNA inserts between them are derived from infectious agents which have previously challenged the bacteria. These inserts generate a complementary RNA which can direct a DNA cutting enzyme, Cas9, to specific base sequences in the DNA of the relevant infectious agents, cutting their DNA and hence inactivating them. Cas9, which means “CRISPR associated protein 9”, was originally studied in Streptococcus pyogenes and is a very large multidomain protein of 158kDa molecular weight. This is inconvenient for many purposes as the DNA needed to express it is correspondingly large, which can present problems with some DNA vector systems. A systematic search for smaller Cas9 homologues showed that Staphylococcus aureus expressed a significantly smaller version, of 124kDa, and we have made a monoclonal antibody to the the C-terminal domain of this, namely MCA-6F7. We also continue to build out our portfolio of antibodies directed against Cnidarian fluorescent proteins, with rabbit antibodies to FP506 (RPCA-FP506) from Zooanthus and EosFP (RPCA-EOSFP) from Lobophyllia hemprichii. Both antibodies are available now. We also release two mouse monoclonals to GFP, MCA-1F1 and MCA-3B11. We add an affinity purified rabbit polyclonal antibody to secretagogin, RPCA-SCGN-ap, to our collection of antibodies to small Calcium binding proteins. These are useful for subclassifying interneurons in the brain. There is a growing interest in the detection of GFAP in blood and CSF as a potential biomarker of CNS damage and disease states. Since the human and rodent proteins have some significant differences at the primary sequence level, we have now expressed full length recombinant rat GFAP, Prot-r-GFAP-rat, which is a more useful protein standard for workers using rodent models. Finally we release an excellent affinity purified antibody to laminin, RPCA-Laminin-ap, which stains blood vessels in the brain beautifully.

October 2015 News

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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 some 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.

July 2015 News

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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 in 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 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, from which were then purified. 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

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We release more new antibodies. To complement our calbindin antibodies we now release a new antibody to calretinin. Calbindin and calretinin are small calcium binding proteins which are each found in subsets of neurons in the CNS, and good antibodies to them are therefore useful for the identification of specific neuronal cell types. The new mouse monoclonal antibody to calretinin is MCA-3G9. We went to the trouble of showing that this antibody does not cross-react with the related proteins calbindin, parvalbumin and secretagogin. 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 antibody MCA-38F3.

New Antibodies and New Data

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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.

New Recombinant Human Proteins and Antibodies

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We have had a long time interest in the intermediate or 10nm filaments of the nervous system, antibodies to which are very useful cell type and developmental markers. There has also been considerable recent interest in the detection of these proteins as biomarkers of different kinds of CNS damage and disease. In the last few weeks we have manufactured large amounts of several of these proteins in recombinant form. These will be used as ELISA protein standards, for monoclonal antibody epitope mapping and for continued production of more antibodies. All of the proteins were made using the pET expression system, and were expressed in and purified from E. coli.  All are expressed from cDNAs encoding the full length human protein sequence, but were codon optimized for efficient E. coli, expression. We currently have neurofilament NF-L (catalog entry Prot-r-NF-L), vimentin (catalog entry Prot-r-Vim), peripherin (catalog entry Prot-r-Peri), GFAP (catalog entry Prot-r-GFAP) and α-internexin (catalog entry Prot-r-a-Int), and are producing other similar constructs on a routine basis, including those related to general neuroscience. We also introduce new monoclonal antibodies to GAP43, MCA-5E8 and MCA-3H14 and a new monoclonal antibody to microtuble associated protein tau MCA-5B10.