October News
We release two high quality mouse monoclonal antibodies MCA-1B63 and MCA-3H25 to the S-tag, a 15 amino acid sequence originally identified as part of the N-terminal sequence of secreted bovine pancreatic ribonuclease by Frederick M. Richards in the 1950s. The S-tag sequence has been incorporated into many expression vectors and so S-tag antibodies are useful for verification of the correct expression of recombinant proteins, and the peptide can also be used as an affinity tag for protein purification. We have also have kinetic data on both antibodies, have shown they specifically bind bovine pancreatic ribonuclease and that they are useful for verifying the correct molecular size of recombinant constructs containing the S-tag sequence.
We previously made a very popular mouse monoclonal antibody to α-synuclein MCA-2A7 and more recently another mouse monoclonal to β-synuclein MCA-6A10. Since α, β and γ-synuclein are somewhat related in protein sequence, it is important to show that MCA-2A7 is specific for α-synuclein and MCA-6A10 is specific for β-synuclein. We have run the essential western blotting experiment and can show that both antibodies are completely specific for the appropriate proteins, see data under the “additional info” tag on the web page for each antibody. Since we have made recombinant full length human β and γ-synuclein proteins available in house we are now also marketing these also, as PROT-r-SNCB and PROT-r-SNCG.
July News
You may have noticed that we recently updated our homepage, as our previous version became unstable for some unknown probably computer code related reason. We hope you like the newer version.
We are glad to report that our recent paper, “TNFα increases tyrosine hydroxylase expression in human monocytes”, an early version of which was posted on the BioRΧiv server has been accepted for publication in NPJ Parkinson’s disease and can now be freely downloaded, from the journal here and also from Pubmed here. The Nature partner journals series, abbreviated NPJ, are a series of online-only, open access journals spun off by the journal Nature, one of the most prestigious and respected in science. This high impact journal is associated with the journal Parkinson’s Foundation and so provides a good venue to contact scientists and clinicians engaged in Parkinson’s disease research. The paper makes use of a sensitive and novel tyrosine hydroxylase ELISA developed using EnCor antibodies MCA-4H2 and RPCA-TH which we are now preparing for commercialization. For a write up of this work published on the University of Florida web site see here.
We also release a novel goat polyclonal antibody to neurofilament NF-H GPCA-NF-H. This protein is heavily expressed in axons and is also released into blood and CSF following axonal compromise providing a useful surrogate marker of axonal damage and degeneration.
Finally we continue to characterize the epitopes for our various monoclonal antibodies. An interesting recent finding it the mapping of one of our neurofilament NF-L monoclonals MCA-1B11. This binds to a highly conserved region of the dimeric α-helical coiled coil region of human NF-L. Much interest has been focused on the use of a neurofilament NF-L assay from Uman Diagnostics to monitor axonal loss following CNS compromise. The NF light Uman assay is based on two mouse monoclonal antibodies, clone 2.1 for NF-L detection and clone 47.3 for NF-L capture (see Norgren et al. 2002). These two clones are also known as UD1 and UD2 respectively. Interestingly our MCA-1B11 binds to the same epitope as the Uman detection reagent, UD1/clone 2.1. We are currently characterizing the epitope for the other Uman reagent and will publish our findings on this. This work will lead to a better understanding of exactly how the Uman assay works and will lead to the development of novel second generation NF-L assays.
April News
We publish another collaboration between scientists at the University of Florida and EnCor, “TNF increases tyrosine hydroxylase expression in human monocytes” by Madison et al., BioRΧiv 10.1101/2021.04.13.439627v1, It describes in detail an ELISA developed using EnCor antibodies which can be used for quantitation of tyrosine hydroxylase (TH) levels in a variety of experimental and clinical contexts. The specific antibodies used are our mouse monoclonal antibody MCA-4H2 and our rabbit polyclonal RPCA-TH. They also used our chicken polyclonal to the cell division marker Ki67 CPCA-Ki67. Previously the same group showed that TH levels were detectable in blood cells of healthy control and Parkinson’s patients and that the levels detected in Parkinson’s patients were significantly higher, a potential very significant finding.
We have been very interested in recent studies of NF-L, neurofilament light, as a biomarker of neuronal damage and degeneration. Much of the recent work has been performed with the excellent Uman NF-Light Assay. The epitopes recognized by the two antibodies used in this assay are not known, and so we have been working to fully characterize them. As part of this work we generated a codon optimized cDNA designed to express amino acids 256-400 of the α-helical rod region of human neurofilament NF-L, specifically the Coil 2a and Coil 2b regions. Our full length recombinant human NF-L protein Prot-r-NF-L is widely used as a protein standard in ELISA, Simoa and other kinds of antibody based assays for NF-L detection, including the Uman assay. Our new truncated product, Prot-r-NF-L-rct, contains the epitopes for both the capture and detect NF-L monoclonal antibodies used in the Uman NF-Light assay and also the epitope for our mouse NF-L monoclonal MCA-1B11. This product may therefore be a more convenient protein standard for use with the Uman and similar NF-L assays, and our new data suggests that MCA-1B11 may form the basis of further NF-L assays.
We also updated our product data for several of our recombinant proteins including mCherry Prot-r-mCherry and GFP Prot-r-acGFP.
February 2021 News
We published a preprint on the BioRΧiv server, observations on the biomarkers pNF-H and UCHL1 in the bodily fluids of patients recovering from a common type of thoracic surgery. This report “A study of the neuronal injury biomarkers pNF-H and UCHL1 in serum, CSF and urine in a cohort of thoracic endovascular aortic repair (TEVAR) patients”, by Beck et al., BioRΧiv 10.1101/2021.03.02.433405v1, in addition to documenting blood, CSF and urine levels of these two proteins in a large cohort of TEVAR patients, also provides important quantitative data on the levels of UCHL1 in healthy control blood samples using multiple antibodies, results with antibody data confirmed by mass spectroscopy. We also document the presence of UCHL1 in certain urine samples. We used novel pNF-H and UCHL1 assays developed for the MSD electrochemiluminescent platform using EnCor antibodies and ran all the several hundred assays in the EnCor lab. We also include epitope mapping and other data on our panel of UCHL1 antibodies including the widely used mouse monoclonal antibody MCA-BH7.
We have updated product data for our recombinant full length human proteins, NF-L, GFAP, vimentin and UCHL1, products PROT-r-NF-L, PROT-r-GFAP, PROT-r-Vim and PROT-r-UCHL1 respectively. We now provide the exact amino acid sequence and composition, isoelectric point, molecular weight, extinction coefficient and other relevant data for each protein. These proteins are widely used as protein standard for ELISA, MSD, Luminex and Simoa type assays, and also as immunogens for antibody production. We will provide this information for all of our recombinant proteins shortly and have already done this for our full length recombinant rat GFAP protein PROT-r-GFAP-rat.
January News
We release novel antibodies to one of the CasΦ, a.k.a. Cas-12j, family of enzymes. These are newly described and exceptionally compact enzymes of great potential utility in gene editing by CRISPR. Recent ecosystem and microbiome DNA sequencing and assembly characterized numerous whole genomes of “huge phages” or megaphages, see Al-Shayeb et al. Nature 578:425-431 (2020). These phages have much larger genomes than those of well studies bacteriophages such as phage-λ and members of this family are apparently ubiquitously found in all microbiomes and ecosystems. Interestingly the genomes may include genes which encode novel CRISPR associated or Cas enzymes. The lab of recent Nobel prize winner Professor Jennifer Doudna further characterized some of these megaphage Cas enzymes and showed that they could be used to perform CRISPR but had the significant advantage that they were much smaller in molecular size than typical Cas9 and Cas12 enzymes such as those from S. pyogenes and S. aureus, see Pausch et al. Science 369:333-337 (2020). The amino acid sequence of several of these enzymes was published and the group studied was named CasΦ or Cas12j. The small relative size of CasΦ enzymes leaves more room for other nucleic acid sequences in AAV and other viral vectors which typically have a limited DNA capacity. We made a mouse monoclonal and a rabbit polyclonal to one of these enzymes, CasΦ-2, MCA-5F95 and RPCA-Cas12J respectively. These can be used verify correct expression of CasΦ-2 protein in CRISPR experiments.
We have also made a novel, high quality and very high titer rabbit polyclonal antibody to microtubule associated protein 2 (MAP2), RPCA-MAP2. Antibodies to MAP2 are excellent markers of neuronal dendrites and perikarya and are also useful in developmental studies. This new antibody was made against recombinant forms of the entire MAP2 projection domain using a mixture of our three human MAP2 projection domain constructs and works very well on tissues of transgenic and wild type rodents at dilutions of 1:10,000. It is therefore a very cost effective reagent for routine neuronal identification in cell cultures and tissue sections.
