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48-602MAG
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ABN1350
Sigma-AldrichAnti-ApoE4 Fragment nApoECF Antibody (Asp172)
Anti-ApoE4 Fragment nApoECF Antibody (Asp172) is an antibody against ApoE4 Fragment nApoECF for use in Immunohistochemistry (Paraffin), Western Blotting, Immunofluorescence.
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Apolipoprotein E (UniProt P02649; also known as Apo-E) is encoded by the APOE (also known as AD2, LDLCQ5, LPG) gene (Gene ID 348) in human. Apolipoprotein E (ApoE) is found in the chylomicron and Intermediate-density lipoprotein (IDLs) and is essential for the normal catabolism of triglyceride-rich lipoprotein constituents. ApoE is initially produced with a signal peptide sequence (a.a. 1-18), the removal of which yields the 299 a.a. mature protein. Three additional natural variants (E2, E3, and E4) exist due to polymorphisms. E4 has an allele frequency of approximately 14 percent and has been implicated in atherosclerosis and various CNS disorders, including Alzheimer's disease (AD). ApoE4 is highly susceptible to proteolysis, which impairs its role in cholesterol transport and beta-amyloid removal in the CNS. Various ApoE4 fragments (14–20 kDa) have been identified in the AD brain. The C-terminal portion of apoE has been implicated in binding to beta-amyloid and is localized to plaques, while the N-terminal domain preferentially localizes within neurofibrillary tangles (NFTs) in the AD brain.
References
Product Information
Format
Affinity Purified
Presentation
Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Anti-ApoE4 Fragment nApoECF Antibody (Asp172) is an antibody against ApoE4 Fragment nApoECF for use in Immunohistochemistry (Paraffin), Western Blotting, Immunofluorescence.
Key Applications
Immunohistochemistry (Paraffin)
Western Blotting
Immunofluorescence
Application Notes
Immunohistochemistry Analysis: A 1:1,000 dilution from a representative lot detected little ApoE4 fragment nApoECF (Asp172) immunoreactivity in normal human brain tissue.
Western Blotting Analysis: 1.0 µg/mL from a representative lot detected ApoE4 fragment nApoECF (Asp172) in 10 µg of human Alzheimer's diseased brain tissue lysate.
Immunofluorescence Analysis: A representative lot detected a strong nApoECF (n-terminal ApoE4 Cleavage Fragment) immunoreactivity co-localized with that of PHF-1 within Pick bodies of area CA1 by dual-fluorescent immunohistochemistry using free-floating hippocampus tissue sections from a Pick's disease patient (Rohn, T.T., et al. (2013). PLoS One. 8(12):e80180).
Immunofluorescence Analysis: A representative lot detected nApoECF (n-terminal ApoE4 Cleavage Fragment) immunoreactivity co-localized with that of cleaved Tau (Asp421; Cat. No. 36-017) within Pick bodies of area CA1 by dual-fluorescent immunohistochemistry using free-floating hippocampus tissue sections from a Pick's disease patient (Rohn, T.T., et al. (2013). PLoS One. 8(12):e80180).
Immunofluorescence Analysis:A representative lot detected the the nApoECF (n-terminal ApoE4 Cleavage Fragment) immunoreactivity co-localized with the PHF-1-positive neurofibrillary tangles (NFTs) in the frontal cortex of Alzheimer's diseased brains by dual-fluorescent immunohistochemistry using formic acid-treated free-floating sections (Rohn, T.T., et al. (2012). Brain Res. 1475:106-115).
Immunohistochemistry Analysis: A representative lot detected a strong nApoECF (n-terminal ApoE4 Cleavage Fragment) immunoreactivity within Pick bodies of area CA1 among 4 out of 5 Pick's disease patients-derived free-floating hippocampus specimens (Rohn, T.T., et al. (2013). PLoS One. 8(12):e80180).
Immunohistochemistry Analysis: A representative lot detected the specific association of the nApoECF (n-terminal ApoE4 Cleavage Fragment) immunoreactivity with the neurofibrillary tangles (NFTs), but not within the Abeta-positive senile plaques, in the frontal cortex of Alzheimer's diseased brains using formic acid-treated free-floating sections (Rohn, T.T., et al. (2012). Brain Res. 1475:106-115).
Western Blotting Analysis: A representative lot detected the 18 kDa nApoECF (n-terminal ApoE4 Cleavage Fragment; a.a. 1-172) present in the preparations of bacterially expressed human ApoE4, but not the full-length ApoE4 itself or the caspse-3-cleaved 16 kDa ApoE4 fragment (Rohn, T.T., et al. (2012). Brain Res. 1475:106-115).
Note: This antibody will detect any ApoE fragments with Asp172 at the C-terminus. This antibody was raised against a hydrophobic immunogen sequence and therefore exhibits high affinity toward hydrophobic membrane surface. Multiple banding pattern and overall high background are expected when using this antibody for Western blotting applications, especially when employing tissue samples.
Biological Information
Immunogen
Linear peptide corresponding to the C-terminal sequence of human ApoE4 fragment nApoECF (Asp172).
Epitope
C-terminus
Concentration
Please refer to lot specific datasheet.
Host
Rabbit
Specificity
Specifically recognizes the ApoE4 N-terminal fragment(s) generated by a proteolytic cleavage between Asp172 and Leu173 of ApoE4. Does not recognize full-length ApoE3 and ApoE4, nor ApoE4 fragments with C-termi other than D172.
Species Reactivity
Human
Species Reactivity Note
Human. Predicted to react with Bovine based on 100% sequence homology.
Evaluated by Immunohistochemistry in Alzheimer's diseased human brain tissue.
Immunohistochemistry Analysis: A 1:250 dilution of this antibody detected ApoE4 fragment nApoECF (Asp172) in Alzheimer's diseased human brain tissue.
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Immunolocalization of an amino-terminal fragment of apolipoprotein E in the Pick's disease brain. Rohn, TT; Day, RJ; Catlin, LW; Brown, RJ; Rajic, AJ; Poon, WW PloS one
8
e80180
2013
Although the risk factor for apolipoprotein E (apoE) polymorphism in Alzheimer's disease (AD) has been well described, the role that apoE plays in other neurodegenerative diseases, including Pick's disease, is not well established. To examine a possible role of apoE in Pick's disease, an immunohistochemical analysis was performed utilizing a novel site-directed antibody that is specific for an amino-terminal fragment of apoE. Application of this antibody, termed the amino-terminal apoE cleavage fragment (nApoECF) antibody, consistently labeled Pick bodies within area CA1 of the hippocampus in 4 of the 5 cases examined. Co-localization of the nApoECF antibody with PHF-1, a general marker for Pick bodies, as well as with an antibody to caspase-cleaved tau (TauC3) was evident within the hippocampus. While staining of the nApoECF antibody was robust in area CA1, little co-localization with PHF-1 in Pick bodies within the dentate gyrus was observed. A quantitative analysis indicated that approximately 86% of the Pick bodies identified in area CA1 labeled with the nApoECF antibody. The presence of truncated apoE within Pick bodies suggests a broader role of apoE beyond AD and raises the question as to whether this protein contributes to pathogenesis associated with Pick's disease.
Identification of an amino-terminal fragment of apolipoprotein E4 that localizes to neurofibrillary tangles of the Alzheimer's disease brain. Rohn, TT; Catlin, LW; Coonse, KG; Habig, JW Brain research
1475
106-15
2011
Although the risk factor for harboring the apolipoprotein E4 (apoE4) allele in late-onset Alzheimer's disease (AD) is well known, the mechanism by which apoE4 contributes to AD pathogenesis has yet to be clarified. Preferential cleavage of the ApoE4 isoform relative to other polymorphic forms appears to be significant, as the resulting fragments are associated with hallmarks of AD. To examine the possible role of apoE4 proteolysis in AD, we designed a site-directed antibody directed at position D172, which would yield a predicted amino-terminal fragment previously identified in AD brain extracts. Western blot analysis utilizing this novel antibody, termed the amino-terminal apoE4 cleavage fragment (nApoE4CF) Ab, consistently identified the predicted amino-terminal fragment (∼18kDa) in several commercially available forms of human recombinant apoE4 purified from E. coli. Mass spectrometry confirmed the identity of this 18kDa fragment as being an amino-terminal fragment of apoE4. Immunohistochemical experiments indicated the nApoE4CF Ab specifically labeled neurofibrillary tangles (NFTs) in AD frontal cortex sections that colocalized with the mature tangle marker PHF-1. Taken together, these results suggest a novel cleavage event of apoE4, generating an amino-terminal fragment that localizes within NFTs of the AD brain.
