Recombinant Anti-Filamin A antibody [EP2405Y] (ab76289)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2405Y] to Filamin A
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Filamin A antibody [EP2405Y]
See all Filamin A primary antibodies -
Description
Rabbit monoclonal [EP2405Y] to Filamin A -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human Filamin A (C terminal). The exact sequence is proprietary.
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Positive control
- COS-1, HeLa, HepG2, 3T3 and C6 cell lysate and human uterus 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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.5% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2405Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab76289 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 |
|---|---|---|
| ICC/IF | Use at an assay dependent concentration. PubMed: 21507248 | |
| WB | 1/250000 - 1/500000. Detects a band of approximately 281 kDa (predicted molecular weight: 281 kDa). | |
| IHC-P | 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. | |
| Flow Cyt | 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Promotes orthogonal branching of actin filaments and links actin filaments to membrane glycoproteins. Anchors various transmembrane proteins to the actin cytoskeleton and serves as a scaffold for a wide range of cytoplasmic signaling proteins. Interaction with FLNA may allow neuroblast migration from the ventricular zone into the cortical plate. Tethers cell surface-localized furin, modulates its rate of internalization and directs its intracellular trafficking. -
Tissue specificity
Ubiquitous. -
Involvement in disease
Defects in FLNA are the cause of periventricular nodular heterotopia type 1 (PVNH1) [MIM:300049]; also called nodular heterotopia, bilateral periventricular (NHBP or BPNH). PVNH is a developmental disorder characterized by the presence of periventricular nodules of cerebral gray matter, resulting from a failure of neurons to migrate normally from the lateral ventricular proliferative zone, where they are formed, to the cerebral cortex. PVNH1 is an X-linked dominant form. Heterozygous females have normal intelligence but suffer from seizures and various manifestations outside the central nervous system, especially related to the vascular system. Hemizygous affected males die in the prenatal or perinatal period.
Defects in FLNA are the cause of periventricular nodular heterotopia type 4 (PVNH4) [MIM:300537]; also known as periventricular heterotopia Ehlers-Danlos variant. PVNH4 is characterized by nodular brain heterotopia, joint hypermobility and development of aortic dilation in early adulthood.
Defects in FLNA are the cause of otopalatodigital syndrome type 1 (OPD1) [MIM:311300]. OPD1 is an X-linked dominant multiple congenital anomalies disease mainly characterized by a generalized skeletal dysplasia, mild mental retardation, hearing loss, cleft palate, and typical facial anomalies. OPD1 belongs to a group of X-linked skeletal dysplasias known as oto-palato-digital syndrome spectrum disorders that also include OPD2, Melnick-Needles syndrome (MNS), and frontometaphyseal dysplasia (FMD). Remodeling of the cytoskeleton is central to the modulation of cell shape and migration. FLNA is a widely expressed protein that regulates re-organization of the actin cytoskeleton by interacting with integrins, transmembrane receptor complexes and second messengers. Males with OPD1 have cleft palate, malformations of the ossicles causing deafness and milder bone and limb defects than those associated with OPD2. Obligate female carriers of mutations causing both OPD1 and OPD2 have variable (often milder) expression of a similar phenotypic spectrum.
Defects in FLNA are the cause of otopalatodigital syndrome type 2 (OPD2) [MIM:304120]; also known as cranioorodigital syndrome. OPD2 is a congenital bone disorder that is characterized by abnormally modeled, bowed bones, small or absent first digits and, more variably, cleft palate, posterior fossa brain anomalies, omphalocele and cardiac defects.
Defects in FLNA are the cause of frontometaphyseal dysplasia (FMD) [MIM:305620]. FMD is a congenital bone disease characterized by supraorbital hyperostosis, deafness and digital anomalies.
Defects in FLNA are the cause of Melnick-Needles syndrome (MNS) [MIM:309350]. MNS is a severe congenital bone disorder characterized by typical facies (exophthalmos, full cheeks, micrognathia and malalignment of teeth), flaring of the metaphyses of long bones, s-like curvature of bones of legs, irregular constrictions in the ribs, and sclerosis of base of skull.
Defects in FLNA are the cause of X-linked congenital idiopathic intestinal pseudoobstruction (CIIPX) [MIM:300048]. CIIPX is characterized by a severe abnormality of gastrointestinal motility due to primary qualitative defects of enteric ganglia and nerve fibers. Affected individuals manifest recurrent signs of intestinal obstruction in the absence of any mechanical lesion.
Defects in FLNA are the cause of FG syndrome type 2 (FGS2) [MIM:300321]. FG syndrome (FGS) is an X-linked disorder characterized by mental retardation, relative macrocephaly, hypotonia and constipation.
Defects in FLNA are the cause of terminal osseous dysplasia (TOD) [MIM:300244]. A rare X-linked dominant male-lethal disease characterized by skeletal dysplasia of the limbs, pigmentary defects of the skin and recurrent digital fibroma during infancy. A significant phenotypic variability is observed in affected females.
Defects in FLNA are the cause of cardiac valvular dysplasia X-linked (CVDX) [MIM:314400]. A rare X-linked heart disease characterized by mitral and/or aortic valve regurgitation. The histologic features include fragmentation of collagenous bundles within the valve fibrosa and accumulation of proteoglycans, which produces excessive valve tissue leading to billowing of the valve leaflets. -
Sequence similarities
Belongs to the filamin family.
Contains 1 actin-binding domain.
Contains 2 CH (calponin-homology) domains.
Contains 24 filamin repeats. -
Domain
Comprised of a NH2-terminal actin-binding domain, 24 internally homologous repeats and two hinge regions. Repeat 24 and the second hinge domain are important for dimer formation. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR (By similarity). Phosphorylation extent changes in response to cell activation.
The N-terminus is blocked. -
Cellular localization
Cytoplasm > cell cortex. Cytoplasm > cytoskeleton. - Information by UniProt
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Database links
- Entrez Gene: 2316 Human
- Entrez Gene: 192176 Mouse
- Entrez Gene: 293860 Rat
- Omim: 300017 Human
- SwissProt: P21333 Human
- SwissProt: Q8BTM8 Mouse
- Unigene: 195464 Human
- Unigene: 295533 Mouse
see all -
Alternative names
- ABP 280 antibody
- ABP-280 antibody
- Actin-binding protein 280 antibody
see all
Images
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![Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)](https://www.abcam.com/ps/products/76/ab76289/Images/ab76289-283574-ab76289GDBKOWB.jpg)
Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: FLNA knockout HAP1 whole cell lysate (20 µg)
Lane 3: HepG2 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab76289 observed at 281 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab76289 was shown to specifically react with FLNA when FLNA knockout samples were used. Wild-type and FLNA knockout samples were subjected to SDS-PAGE. Ab76289 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 250000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)](https://www.abcam.com/ps/products/76/ab76289/Images/ab76289-266069-ab76289wb.jpg)
Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)All lanes : Anti-Filamin A antibody [EP2405Y] (ab76289) at 1/10000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 2 : COS-1 (Cercopithecus aethiops kidney) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/1000 dilution
Predicted band size: 281 kDa
Additional bands at: 281 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 secondsBlocking buffer: 5% NFDM/TBST
Diluting buffer: 5% NFDM/TBST
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![Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)](https://www.abcam.com/ps/products/76/ab76289/Images/ab76289-269567-ab76289FC1.jpg)
Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)ab76289 staining Filamin A in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin A antibody [EP2405Y] (ab76289)](https://www.abcam.com/ps/products/76/ab76289/Images/ab76289_2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin A antibody [EP2405Y] (ab76289)Immunohistochemical analysis of paraffin-embedded human uterus using ab76289 at a 1/100 dilution.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-Filamin A antibody [EP2405Y] (ab76289)](https://www.abcam.com/ps/products/76/ab76289/Images/ab76289_3.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Filamin A antibody [EP2405Y] (ab76289)Immunofluorescent staining of HeLa cells using ab76289 at a 1/100 dilution. -
![Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)](https://www.abcam.com/ps/products/76/ab76289/Images/Filamin-A-Primary-antibodies-ab76289-3.jpg)
Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)Overlay histogram showing A431 cells stained with ab76289 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76289 , 1/100 dilutio) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions. -
![Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)](https://www.abcam.com/ps/products/76/ab76289/Images/ab76289_1.jpg)
Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)All lanes : Anti-Filamin A antibody [EP2405Y] (ab76289) at 1/500000 dilution
Lane 1 : COS-1 (African green monkey kidney fibroblast-like cell line) cell lysate
Lane 2 : Hela cell lysate
Lane 3 : 3T3 cell lysate
Lane 4 : C6 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/1000 dilution
Predicted band size: 281 kDa
Observed band size: 281 kDa
Protocols
References (21)
ab76289 has been referenced in 21 publications.
- Forrester A et al. A selective ER-phagy exerts procollagen quality control via a Calnexin-FAM134B complex. EMBO J 38:N/A (2019). PubMed: 30559329
- Coelho MCA et al. Clinical significance of filamin A in patients with acromegaly and its association with somatostatin and dopamine receptor profiles. Sci Rep 9:1122 (2019). PubMed: 30718563
- Vähätupa M et al. SWATH-MS Proteomic Analysis of Oxygen-Induced Retinopathy Reveals Novel Potential Therapeutic Targets. Invest Ophthalmol Vis Sci 59:3294-3306 (2018). PubMed: 30025079
- Latham SL et al. Variants in exons 5 and 6 of ACTB cause syndromic thrombocytopenia. Nat Commun 9:4250 (2018). PubMed: 30315159
- Krawczyk KM et al. Injury induced expression of caveolar proteins in human kidney tubules - role of megakaryoblastic leukemia 1. BMC Nephrol 18:320 (2017). PubMed: 29065889
