Anti-Filamin A 抗体 [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 (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human, African green monkey
Related conjugates and formulations
製品の概要
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製品名
Anti-Filamin A antibody [EP2405Y]
Filamin A 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP2405Y] to Filamin A -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, WB, IHC-P, Flow Cyt (Intra)more details
適用なし: IP -
種交差性
交差種: Mouse, Rat, Human, African green monkey -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- COS-1, HeLa, HepG2, 3T3 and C6 cell lysate and human uterus tissue.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP2405Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab76289の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF |
1/100.
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WB | (4) |
1/250000 - 1/500000. Detects a band of approximately 281 kDa (predicted molecular weight: 281 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
特記事項 |
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ICC/IF
1/100. |
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 (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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機能
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. -
組織特異性
Ubiquitous. -
関連疾患
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. -
配列類似性
Belongs to the filamin family.
Contains 1 actin-binding domain.
Contains 2 CH (calponin-homology) domains.
Contains 24 filamin repeats. -
ドメイン
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. -
翻訳後修飾
Phosphorylated upon DNA damage, probably by ATM or ATR (By similarity). Phosphorylation extent changes in response to cell activation.
The N-terminus is blocked. -
細胞内局在
Cytoplasm > cell cortex. Cytoplasm > cytoskeleton. - Information by UniProt
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参照データベース
- 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
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別名
- ABP 280 antibody
- ABP-280 antibody
- Actin-binding protein 280 antibody
see all
画像
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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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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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ab76289 staining Filamin Ain the human cell line HeLa (human cervix adenocarcinoma) by intracellular 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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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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Immunofluorescent staining of HeLa cells using ab76289 at a 1/100 dilution.
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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.
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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
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (36)
ab76289 は 36 報の論文で使用されています。
- Bertović I et al. Early Endosomal GTPase Rab5 (Ras-Related Protein in Brain 5) Regulates GPIbβ (Glycoprotein Ib Subunit β) Trafficking and Platelet Production In Vitro. Arterioscler Thromb Vasc Biol 42:e10-e26 (2022). PubMed: 34732055
- Liu L et al. Myocardin regulates exon usage in smooth muscle cells through induction of splicing regulatory factors. Cell Mol Life Sci 79:459 (2022). PubMed: 35913515
- Schink KO et al. The phosphoinositide coincidence detector Phafin2 promotes macropinocytosis by coordinating actin organisation at forming macropinosomes. Nat Commun 12:6577 (2021). PubMed: 34772942
- Muniappan L et al. Inducible Depletion of Calpain-2 Mitigates Abdominal Aortic Aneurysm in Mice. Arterioscler Thromb Vasc Biol 41:1694-1709 (2021). PubMed: 33761765
- Chen N et al. Hippocampus chronic deep brain stimulation induces reversible transcript changes in a macaque model of mesial temporal lobe epilepsy. Chin Med J (Engl) 134:1845-1854 (2021). PubMed: 34267068