References

General Antibodies

We have generated several monoclonal antibodies specific for short protein tags (e.g. myc-tag), widely used fusion protein moieties (e.g. glutathione-S-transferase [GST] or green fluorescent protein [GFP]), or commonly used marker proteins (e.g. lamin A/C). Some of these antibodies can be purchased from our licensing partners: anti myc-tag (clone 4A6) from Millipore, anti lamin A/C (clone 4C11) from Cell Signaling, ActiveMotif, Enzo Life Sciences. In addition, the MFPL Monoclonal Antibody Facility also makes them directly and exclusively available for MFPL members. Please contact the facility manager for details. To see a list of currently available antibodies please follow the link below.

Mutant/SNP specific Antibodies

Missense mutations or single nucleotide polymorphisms (SNP) can alter the function of a protein with fatal consequences for the affected individual. How a single amino acid substitution in a protein affects its properties is difficult to study in the context of the cellular proteome, because mutant proteins can often not be traced in cells due to the lack of mutation-specific detection tools. Antibodies, however, with their exquisite epitope specificity, permit the detection of single amino acid substitutions but are not available for the vast majority of disease-causing mutant proteins. Hence, one of our goals is the development of monoclonal antibodies specific for disease-linked point-mutant proteins, e.g. human A-type lamins or apolipoprotein E isoforms.

ApoE4

Human apolipoprotein E (ApoE) is a soluble apolipoprotein. It interacts with low-density lipoprotein receptors, thereby playing a pivotal role in the transport of cholesterol and other lipids in the plasma as well as the central nervous system (1). In humans, three polymorphic forms of ApoE are found, which differ from each other at two amino acid positions (see Figure a). ApoE3 is the most prevalent and considered the “wild-type” isoform. ApoE2 is the least common isoform (7-8%). The ApoE4 isoform has been identified as a major genetic risk factor for the development of late-onset Alzheimer´s disease (AD), and a number of potential mechanisms have been suggested for how ApoE4 could mediate this dramatically elevated risk of acquiring AD. In addition, the epsilon-4 allele has also been linked to other neurological disorders, atherosclerosis, or multiple sclerosis (1). We have generated a monoclonal antibody specific for the ApoE4 isoform, which can be used in a variety of techniques, including Western blot, immunoprecipitation and immunofluorescence.

(1) Apolipoprotein E and its receptors in Alzheimer´s disease: pathways, pathogenesis and therapy. Bu G. Nat Rev Neuroscience. 2009

Lamin A R453W R482W

Laminopathies represent a group of rare inherited human diseases, which are caused by mutations in genes encoding components of the nuclear lamina, in particular the LMNA gene coding for lamin A and lamin C. Laminopathies show a wide variety of clinical symptoms ranging from skeletal muscle dystrophies and cardiomyopathies to lipodystrophies and premature aging syndromes. Most mutations in the LMNA gene are single point missense mutations, which are inherited in an autosomal dominant fashion, resulting in the contemporaneous expression of wild-type and mutant lamin A/C in dieseased cells. Hence, to study the effects of lamin A/C point-mutants scientists have so far had to rely on over-expression studies or knock-out or transgenic animal models. We have recently generated and characterized two mouse monoclonal antibodies specific for the disease-associated lamin A/C point-mutants, R453W and R482W, respectively (1). These mutant-specific antibodies will open up new avenues to investigate the molecular basis of laminopathies, and the generation of similar mutant-specific antibodies will provide a way to study new aspects of many hereditary or acquired somatic diseases in primary human cells.

(1) Monoclonal Antibodies Specific for Disease-Associated Point-Mutants: Lamin A/C R453W and R482W. Roblek M, Schüchner S et al. PLoS ONE. 2010

Progerin

We have generated the only commercially available antibody against human progerin. Progerin is a truncated version of lamin A, causing Hutchinson-Gilford progeria syndrome (HGPS), a premature aging syndrome (1). Approximately 80% of HGPS-affected individuals are heterozygous for a silent point mutation within exon 11 of the LMNA gene (C1824T). This mutation activates a cryptic splice site and gives rise to a form of lamin A with a deletion of 50 amino acids near the C-terminus, resulting in aberrant post-translational processing and localization of progerin. Interestingly, expression of progerin by sporadic use of the cryptic splice site has also been observed in cells from healthy individuals, suggesting a possible role in physiological aging (2, 3).

(1) Human laminopathies: nuclei gone genetically awry. Capell BC, Collins FS. Nat Rev Genet. 2006

(2) Lamin A-dependent nuclear defects in human aging. Scaffidi P, Misteli T. Science. 2006

(3) Lamin A-dependent misregulation of adult stem cells associated with accelerated ageing. Scaffidi P, Misteli T. Nat Cell Biol. 2008

Modification specific Antibodies

Post-translational modifications of proteins are key components that contribute to the diversity of the proteome, affecting e.g. protein structure, function, enzymatic activity, localization, or stability. While protein phosphorylation is considered the most common posttranslational modification, more than 200 different covalently attached modifications targeting most proteinogenic amino acids are known today (1). Prime examples of how post-translational modifications can alter a protein´s function are histones, which are modified by a variety of moieties (e.g. acetylation, methylation, phosphorylation, ubiquitination) on a number of N-terminal amino acids. Histone modifications are highly dynamic and are crucial for the regulation of gene transcription, replication, recombination and DNA repair. In the course of an FWF-funded project we have generated a series of antibodies specifically recognizing methylated lysine residues in histone H3.

(1) Posttranslational modification of proteins. Walsh CT. 2006

Custom Antibodies

Since its establishment in May 2009, the MFPL Monoclonal Antibody Facility has attracted both MFPL-internal as well as external researchers, who have sought to develop customized monoclonal antibodies against a variety of antigens, including: Karl Kuchler (MFPL), Graham Warren (MFPL), David Virshup (IMB Singapore), Pavel Kovarik (MFPL), Peggy Stolt-Bergner (IMP), or Harald Sitte (Med. Univ. Wien).

 

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