Recombinant Anti-FE65 antibody [EPR3538] (ab91650)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3538] to FE65
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-FE65 antibody [EPR3538]
See all FE65 primary antibodies -
Description
Rabbit monoclonal [EPR3538] to FE65 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human FE65 aa 650 to the C-terminus. The exact sequence is proprietary.
Database link: O00213 -
Positive control
- SH-SY5Y cell lysate and Human brain tissue
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3538 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab91650 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/2000 - 1/5000. Predicted molecular weight: 77 kDa.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
The use of an HRP/AP polymerized antibody is recommended. We have compared both the HRP-conjugated and the polymerized HRP and found stronger signals can be obtained with the polymerized antibody. |
Notes |
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WB
1/2000 - 1/5000. Predicted molecular weight: 77 kDa. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. The use of an HRP/AP polymerized antibody is recommended. We have compared both the HRP-conjugated and the polymerized HRP and found stronger signals can be obtained with the polymerized antibody. |
Target
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Function
Transcription coregulator that can have both coactivator and corepressor functions. Adapter protein that forms a transcriptionally active complex with the gamma-secretase-derived amyloid precursor protein (APP) intracellular domain. Plays a central role in the response to DNA damage by translocating to the nucleus and inducing apoptosis. May act by specifically recognizing and binding histone H2AX phosphorylated on 'Tyr-142' (H2AXY142ph) at double-strand breaks (DSBs), recruiting other pro-apoptosis factors such as MAPK8/JNK1. Required for histone H4 acetylation at double-strand breaks (DSBs). Its ability to specifically bind modified histones and chromatin modifying enzymes such as KAT5/TIP60, probably explains its trancription activation activity. Function in association with TSHZ3, SET and HDAC factors as a transcriptional repressor, that inhibits the expression of CASP4. Associates with chromatin in a region surrounding the CASP4 transcriptional start site(s). -
Tissue specificity
Highly expressed in brain; strongly reduced in post-mortem elderly subjects with Alzheimer disease. -
Sequence similarities
Contains 2 PID domains.
Contains 1 WW domain. -
Post-translational
modificationsPhosphorylated following nuclear translocation. Phosphorylation at Tyr-546 enhances the transcription activation activity and reduces the affinity with RASD1/DEXRAS1. -
Cellular localization
Cell membrane. Cytoplasm. Nucleus. Cell projection > growth cone. Colocalizes with TSHZ3 in axonal growth cone (By similarity). In normal conditions, it mainly localizes to the cytoplasm, while a small fraction is tethered to the cell membrane via its interaction with APP. Following exposure to DNA damaging agents, it is released from cell membrane and translocates to the nucleus. Nuclear translocation is under the regulation of APP. Colocalizes with TSHZ3 in the nucleus. - Information by UniProt
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Database links
- Entrez Gene: 322 Human
- Omim: 602709 Human
- SwissProt: O00213 Human
- Unigene: 372840 Human
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Alternative names
- Adaptor protein FE65a2 antibody
- Amyloid beta (A4) precursor protein binding family B member 1 antibody
- Amyloid beta A4 precursor protein binding family B antibody
see all
Images
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All lanes : Anti-FE65 antibody [EPR3538] (ab91650) at 1/2000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : APBB1 knockout HEK-293T cell lysate
Lane 3 : SH-SY5Y cell lysate
Lane 4 : A431 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 77 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-FE65 antibody [EPR3538] staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab91650 was shown to bind specifically to FE65. A band was observed at 65/85 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in APBB1 knockout cell line ab267294 (knockout cell lysate ab257833). To generate this image, wild-type and APBB1 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Anti-FE65 antibody [EPR3538] (ab91650) at 1/2000 dilution + SH-SY5Y cell lysates at 10 µg
Secondary
HRP labelled goat anti-rabbit IgG at 1/1000 dilution
Predicted band size: 77 kDa -
ab91650, at 1/50 dilution, staining FE65 in formalin-fixed, paraffin-embedded Human brain tissue, by Immunohistochemistry.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab91650 has been referenced in 4 publications.
- Xu P et al. GSAP regulates lipid homeostasis and mitochondrial function associated with Alzheimer's disease. J Exp Med 218:N/A (2021). PubMed: 34156424
- Iannuzzi F et al. Fyn Tyrosine Kinase Elicits Amyloid Precursor Protein Tyr682 Phosphorylation in Neurons from Alzheimer's Disease Patients. Cells 9:N/A (2020). PubMed: 32751526
- Andrew RJ et al. Quantitative interaction proteomics reveals differences in the interactomes of amyloid precursor protein isoforms. J Neurochem 149:399-412 (2019). PubMed: 30664241
- Langlands H et al. Fe65 Is Phosphorylated on Ser289 after UV-Induced DNA Damage. PLoS One 11:e0155056 (2016). PubMed: 27176072