Overview

  • Product name

    Anti-Filamin A (phospho S2152) + Filamin B (phospho S2107) antibody [EP2310AY]
  • Description

    Rabbit monoclonal [EP2310AY] to Filamin A (phospho S2152) + Filamin B (phospho S2107)
  • Host species

    Rabbit
  • Specificity

    ab75978 detects Filamin A phosphorylated on serine 2152 and Filamin B phosphorylated on serine 2107.
  • Tested applications

    Suitable for: ICC/IF, ELISA, WB, IHC-Pmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Filamin A (phospho S2152) + Filamin B (phospho S2107) aa 2100-2200.
    Database link: P21333

  • Positive control

    • HeLa cell lysates, untreated or treated with calyculin A; human uterus tissue.
  • General notes

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 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

    EP2310AY
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab75978 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 1/100 - 1/200.
ELISA Use at an assay dependent concentration. PubMed: 21507248
WB 1/10000 - 1/20000. Detects a band of approximately 281 kDa (predicted molecular weight: 281 kDa).
IHC-P 1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Relevance

      Filamin A-P21333. 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. Involved in ciliogenesis. Filamin B-O75369. Function: Connects cell membrane constituents to the actin cytoskeleton. May promote orthogonal branching of actin filaments and links actin filaments to membrane glycoproteins. Anchors various transmembrane proteins to the actin cytoskeleton. Interaction with FLNA may allow neuroblast migration from the ventricular zone into the cortical plate. Various interactions and localizations of isoforms affect myotube morphology and myogenesis. Isoform 6 accelerates muscle differentiation in vitro.
    • Database links

    • Alternative names

      • ABP-278 antibody
      • ABP-280 antibody
      • ABP-280 homolog antibody
      • Actin-binding protein 280 antibody
      • Actin-binding-like protein antibody
      • Alpha-filamin antibody
      • Beta-filamin antibody
      • Endothelial actin-binding protein antibody
      • Fh1 antibody
      • Filamin homolog 1 antibody
      • Filamin-1 antibody
      • Filamin-3 antibody
      • Filamin-A antibody
      • Filamin-B antibody
      • FLN antibody
      • FLN-A antibody
      • FLN1 antibody
      • FLN1L antibody
      • FLN3 antibody
      • FLNA antibody
      • FLNB antibody
      • Non-muscle filamin antibody
      • TABP antibody
      • TAP antibody
      • Thyroid autoantigen antibody
      • Truncated ABP antibody
      • Truncated actin-binding protein antibody
      see all

    Images

    • All lanes : Anti-Filamin A (phospho S2152) + Filamin B (phospho S2107) antibody [EP2310AY] (ab75978) at 1/20000 dilution

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 5nM calyculin A for 2 hours whole cell lysate with 5% NFDM/TBST
      Lane 2 : Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 281 kDa
      Observed band size: 281 kDa


      Exposure time: 180 seconds
    • Immunofluorescence analysis of U. parvum infected BPH-1 (Human benign prostate) cells staining Filamin A (phospho S2152) and Filamin B (phospho S2107) with ab75978 (red). Cells were fixed with formaldehyde before incubating with primary antibody. An AlexaFluor®594-conjugated goat anti-rabbit IgG was used as the secondary antibody.

    • ab75978 at 1/100 dilution staining Filamin A (phospho S2152) and Filamin B (phospho S2107) in human uterus by Immunohistochemistry, Paraffin-embedded tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Allam AB  et al. Ureaplasma parvum infection alters filamin A dynamics in host cells. BMC Infect Dis 11:101 (2011). ICC/IF, ELISA ; Rat . Read more (PubMed: 21507248) »
    • Bogatcheva NV  et al. Molecular mechanisms mediating protective effect of cAMP on lipopolysaccharide (LPS)-induced human lung microvascular endothelial cells (HLMVEC) hyperpermeability. J Cell Physiol 221:750-9 (2009). WB ; Human . Read more (PubMed: 19725051) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    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

    Answer

    Thank you for your inquiry. HeLa cells are serum-starved overnight, and then treated with 100 nM Calyculin A for 30 min at 37 deg C. Calyculin A was chosen because it is a known serine/threonine phosphatase inhibitor. 1.Resjö, S. et al. (1999) Biochem J 341 ( Pt 3), 839-45. 2.Ishihara, H. et al. (1989) Biochem Biophys Res Commun 159, 871-7. I hope this information helps. Please let me know if you have questions.

    Read More

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