The Immunogen: TDP43 was originally identified as a protein which binds to the "transactivation response" (TAR) sequence found in the long terminal repeat of the HIV-1 virus genome (1). UV cross-linking of HeLa cell extract revealed a 43kDa protein which was cloned and sequenced and shown to contain a two copies of the ~90 amino acid RRM domain. RRM is an acronym for RNA Recognition Motif, and this domain is found in many proteins which bind single stranded RNA and some which bind single stranded DNA. The protein is frequently refered to by the acronym TDP43 corresponds to "TAR DNA binding protein of molecular weight 43kDa", although the Human Genome Organization (HUGO) Gene Nomenclature Committee (HGNC) name is TARDBP. Northern blots showed that the protein is ubiquitous in tissue expression. Much interest has been focused on TDP43 recently due to its association with the inclusions seen in frontotemporal lobar degeneration and Amyotrophic Lateral Sclerosis (2). The protein is present in these inclusions in a partially degraded, hyperphosphorylated and ubiquitinated form. For a recent review of the role in TDP43 in various neurological disorders see reference 3.

Figures: Left: Blots of MCA-3H8 on crude extract of mouse brain nuclear fraction (left lane) and cytoplasmic fraction (right lane) blotted with monoclonal antibody MCA-3H8. There is a strong clear band in the nuclear preparation running at 43kDa, and a much more minor band in the cytoplasmic fraction. Image courtesy of Irina Madorsky. Right: MCA-3H8 was used to stain a section of formalin fixed adult rat brain, specifically the hippocampus. Hippocampal neuron nuclei are stained strongly. Chicken antibody to GFAP CPCA-GFAP (green) shows the processes of astrocytic glial cells. Nuclei of all cells are revealed with DAPI DNA stain (blue). The TARDP antibody stains neuronal nuclei strongly and the nuclei of some non-neuronal cells much more weakly. Neuronal nuclei therefore look crimson, since they are both red due to the content of TDP43 and blue due to their content of DNA, stained blue with DAPI.

Suggestions for use: Try at dilutions of 1:1,000 and higher for immunofluorescence, 1:5,000 and lower for western blotting.

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References:

1. Ou SH, Wu F, Harrich D, García-Martínez LF and Gaynor RB. Cloning and characterization of a novel cellular protein, TDP-43, that binds to human immunodeficiency virus type 1 TAR DNA sequence motifs. J Virol. 69:3584-96 (1995)..

2. Neumann, M.; Sampathu, D. M.; Kwong, L. K.; Truax, A. C.; Micsenyi, M. C.; Chou, T. T.; Bruce, J.; Schuck, T.; Grossman, M.; Clark, C. M.; McCluskey, L. F.; Miller, B. L.; Masliah, E.; Mackenzie, I. R.; Feldman, H.; Feiden, W.; Kretzschmar, H. A.; Trojanowski, J. Q.; Lee, V. M.-Y. Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Science 314:130-133 (2006).

3. Forman MS, Trojanowski JQ and Lee VM-Y. TDP-43: a novel neurodegenerative proteinopathy. Current Opinions in Neurobiology 17:548-55 (2007).

Antibody Characteristics: This antibody was raised against recombinant full length human his-tagged TDP43 which was expressed in E. coli and purifed by nickel affinity. The antibody is currently known to stain human and rodent TDP43, but likely works on many other species since TDP43 is highly conserved in primary sequence.

Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.

Availability and Price: Available for shipping now, $200 US per aliquot of 100 microliters of mouse ascites fluid, enough for hundreds of experiments. For order form press here

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