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. It also has a single Zinc Finger domain of the ZnF_RBZ subtype found in Ran binding proteins. Ran is a small G protein related to p21-Ras which regulates the import and export of proteins to the nucleus. 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. This antibody has been used in a recent peer reviewed publication, showing that it works well on western blots and on histological sections (4). Data showing that the epitope for this antibody is in the first 150 amino acids of human TDP43, and further images of histological staining is shown here. The HGNC name for this protein is TARDBP.

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.

Antibody characteristics: MCA-3H8 is a mouse IgG1 class antibody and was raised against full length recombinant human TDP43 expressed in and purified from E. coli. MCA-3H8 is known to react with TDP43 from human, cow, pig, mouse, rat and other mammals. Since the TDP43 protein sequence is highly conserved across species barriers, it is likely that the antibody is effective on many other species also.

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

OMIM link: press here

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).

4. Wilson AC, Dugger BN, Dickson DW and Wang DS. TDP-43 in aging and Alzheimer's disease - a review. Int J Clin Exp Pathol. 4:147-55 (2011).

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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