Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Filamin A antibody [EP2405Y] (ab76289)

Reviews (5)Q&A (3)References (21)

Overview

  • 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.

  • Positive control

    • COS-1, HeLa, HepG2, 3T3 and C6 cell lysate and human uterus tissue.

  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • 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

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.
  • Application notes
    Is unsuitable for IP.

    • Target

    • 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
      modifications

      Phosphorylated 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

    • Database links

      see all

    • Alternative names

      • ABP 280 antibody
      • ABP-280 antibody
      • Actin-binding protein 280 antibody
      • Alpha filamin antibody
      • Alpha-filamin antibody
      • APBX antibody
      • CSBS antibody
      • CVD1 antibody
      • Endothelial actin binding protein antibody
      • Endothelial actin-binding protein antibody
      • Filamin 1 antibody
      • Filamin A alpha antibody
      • Filamin A antibody
      • Filamin-1 antibody
      • Filamin-A antibody
      • FLN antibody
      • FLN-A antibody
      • FLN1 antibody
      • FLNA antibody
      • FLNA_HUMAN antibody
      • FMD antibody
      • MNS antibody
      • NHBP antibody
      • Non muscle filamin antibody
      • Non-muscle filamin antibody
      • OPD antibody
      • OPD1 antibody
      • OPD2 antibody
      • XLVD antibody
      • XMVD antibody
      see all

    Images

    • Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)
      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.

    • Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)
      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 seconds


      Blocking buffer: 5% NFDM/TBST

      Diluting buffer: 5% NFDM/TBST

    • Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)
      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)

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin A antibody [EP2405Y] (ab76289)
      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.
    • Immunocytochemistry/ Immunofluorescence - Anti-Filamin A antibody [EP2405Y] (ab76289)
      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)
      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)
      Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)
      All lanes : Anti-Filamin A antibody [EP2405Y] (ab76289) at 1/500000 dilution

      Lane 1 : COS-1 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

    References

    This product has been referenced in:

    • Forrester A  et al. A selective ER-phagy exerts procollagen quality control via a Calnexin-FAM134B complex. EMBO J 38:N/A (2019). Read more (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). Read more (PubMed: 30718563) »
    See all 21 Publications for this product

    Publishing research using ab76289? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-8 of 8 Abreviews or Q&A

    Western blot abreview for Anti-Filamin A antibody [EP2405Y]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (brain extract)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    brain extract
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 11 2016

    Immunohistochemistry (Frozen sections) abreview for Anti-Filamin A antibody [EP2405Y]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Embryo)
    Permeabilization
    Yes - 0.3% TritonX-100, 15 min
    Specification
    Embryo
    Blocking step
    NDS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Fixative
    Methanol/Acetone
    Read More

    Abir Yamak

    Verified customer

    Submitted Sep 05 2016

    Immunohistochemistry (Frozen sections) abreview for Anti-Filamin A antibody [EP2405Y]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Xenopus laevis Tissue sections (Xenopus froglet ventricle)
    Permeabilization
    No
    Specification
    Xenopus froglet ventricle
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 29 2016

    Question

    Product code: 76289
    Inquiry: I am looking for an antibody against the C-terminus of filamin-A but I need to know the exact location where the antibody binds. What is the epitope/immunogen sequence for the rabbit anti-filamin-A antibody EP2405Y (ab76289)? Thanks.

    Read More
    Answer

    Thank you for contacting us and your interest in our products.

    The exact immunogen sequence used to raise the anti-Filamin A antibody [EP2405Y] - Carboxyterminal end (ab76289) is considered proprietary information. However, I can share with you that the synthetic peptide sequence was taken from within the last 100 residues of the human Filamin A (between 2547-2647). I hope this information has been of help.

    If this is not sufficient for your needs, please could you let me know why you need the exact sequence and I can see if I can help you further.

    Otherwise, I wish you all the best with your research.

    Read More

    Question

    Please provide immunogen sequences for the following.
    ab76289
    ab80837
    ab75978
    ab124406
    Using in Indirect ELISA. Please provide list of suitable secondary antibodies, for colorimetric and fluorescent detection.

    Read More
    Answer

    Thank you for your recent telephone enquiry and for your patience. It has taken some time to obtain the immunogen information for you.

    For the two rabbit monoclonal antibodies:

    ab76289
    Immunogen: from within amino acids range:########

    ab75978
    Immunogen: from within amino acids range: #########
    For colorimetric detection, we have the following anti-rabbit HRP conjugated secondary antibodies, tested in ELISA:

    ab6829 Chicken polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP)
    HRP conjugated
    Tested in: IHC-P, WB, ELISA, IM, ICC/IF, Dot, IHC-Fr
    https://www.abcam.com/index.html?datasheet=6829 (or use the following: https://www.abcam.com/index.html?datasheet=6829).

    ab6802 Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP)
    HRP conjugated
    Tested in: WB, ELISA, Immunomicroscopy, Dot Blot, ICC/IF, IHC-P, IHC-Fr
    https://www.abcam.com/index.html?datasheet=6802 (or use the following: https://www.abcam.com/index.html?datasheet=6802).

    Anti-rabbit secondary antibodies for fluorescent detection in ELISA:

    ab6825 Chicken anti-Rabbit IgG H&L (FITC) secondary antibody
    FITC conjugated
    Tested in: Flow Cyt, IHC-P, ELISA, IM, ICC/IF, IHC-Fr
    https://www.abcam.com/index.html?datasheet=6825 (or use the following: https://www.abcam.com/index.html?datasheet=6825).

    ab6798 Donkey anti-Rabbit IgG H&L (FITC) secondary antibody
    FITC conjugated
    Tested in: Immunomicroscopy, Flow Cyt, ICC/IF, IHC-P, IHC-Fr, ELISA
    https://www.abcam.com/index.html?datasheet=6798 (or use the following: https://www.abcam.com/index.html?datasheet=6798).


    For the two mouse antibodies:
    ab80837
    Immunogen: Tissue / cell preparation - platelet filamin as stated on the datasheet. I am sorry the exact epitope will not have been mapped.

    ab124406
    Immunogen: Recombinant full length protein, corresponding to amino acids 1-839 of Human Filamin A with Propietary tag (AAH14654), as stated on the datasheet. I am sorry the exact epitope will not have been mapped.

    Suitable secondaries for Colorimetric detection in ELISA:

    ab112458 Chicken polyclonal Secondary Antibody to Mouse IgG - H&L (HRP)
    HRP conjugated
    Tested in: IHC-P, WB, ELISA, ICC/IF
    https://www.abcam.com/index.html?datasheet=112458 (or use the following: https://www.abcam.com/index.html?datasheet=112458).

    ab6820 Donkey polyclonal Secondary Antibody to Mouse IgG - H&L (HRP)
    HRP conjugated
    Tested in: Dot Blot, ELISA, IHC-P, IHC-Fr, WB, ICC/IF
    https://www.abcam.com/index.html?datasheet=6820 (or use the following: https://www.abcam.com/index.html?datasheet=6820).

    Fluorescent conjugated anti mouse IgG antibodies tested in ELISA

    ab5758 Rabbit F(ab')2 polyclonal Secondary Antibody to Mouse IgG - H&L (FITC), pre-adsorbed
    FITC conjugated
    Tested in:: ICC/IF, Immunomicroscopy, Flow Cyt, ELISA
    https://www.abcam.com/index.html?datasheet=5758 (or use the following: https://www.abcam.com/index.html?datasheet=5758).


    ab5765 Goat F(ab')2 polyclonal Secondary Antibody to Mouse IgG - H&L (FITC), pre-adsorbed
    FITC conjugated
    Tested in: ICC/IF, Immunomicroscopy, Flow Cyt, ELISA
    https://www.abcam.com/index.html?datasheet=5765 (or use the following: https://www.abcam.com/index.html?datasheet=5765).

    I can recommend to review the individual online datasheets for further information before purchasing.

    For the fluorescent conjugation, I have selected above some FITC conjugated antibodies. Please do let me known if you would like to use a different conjugate, and I will be pleased to provide some more suggestions to you.

    As discussed on the telephone, please note that the primary antibodies are tested in indirect ELISA only. I am sorry they have not been tested in sandwich ELISA.

    I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

    Read More

    Question

    is immunogen within aa1522-2283?

    Read More
    Answer

    Thank you for contacting us. The immunogen is not contained within aa 1522-2283.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Western blot abreview for Anti-Filamin A antibody [EP2405Y] - Carboxyterminal end

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (platelet)
    Loading amount
    25 µg
    Specification
    platelet
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 20 minute(s) · Concentration: 3% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 22 2011

    Western blot abreview for Anti-Filamin A antibody [EP2405Y] - Carboxyterminal end

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Hela (human) and 3t3 (mouse))
    Loading amount
    154 µg
    Specification
    Hela (human) and 3t3 (mouse)
    Gel Running Conditions
    Reduced Denaturing (8% sds page)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
    Read More

    Abcam user community

    Verified customer

    Submitted May 15 2009

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