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Anti-Emerin antibody (ab40688)

Key features and details

  • Rabbit polyclonal to Emerin
  • Suitable for: IP, ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Emerin antibody
    See all Emerin primary antibodies

  • Description

    Rabbit polyclonal to Emerin

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Cow

  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human Emerin.

    Read Abcam's proprietary immunogen policy (Peptide available as ab40723)

  • Positive control

    • WB: HEK-293T, HAP1 and HeLa whole cell lysates; HeLa nuclear lysate. IHC-P: HeLa cells.

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 or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab40688 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
IP Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 25, 29 kDa).

Target

  • Function

    Stabilizes and promotes the formation of a nuclear actin cortical network. Stimulates actin polymerization in vitro by binding and stabilizing the pointed end of growing filaments. Inhibits beta-catenin activity by preventing its accumulation in the nucleus. Acts by influencing the nuclear accumulation of beta-catenin through a CRM1-dependent export pathway. Links centrosomes to the nuclear envelope via a microtubule association. EMD and BAF are cooperative cofactors of HIV-1 infection. Association of EMD with the viral DNA requires the presence of BAF and viral integrase. The association of viral DNA with chromatin requires the presence of BAF and EMD. Required for proper localization of non-farnesylated prelamin-A/C.

  • Tissue specificity

    Skeletal muscle, heart, colon, testis, ovary and pancreas.

  • Involvement in disease

    Defects in EMD are the cause of Emery-Dreifuss muscular dystrophy type 1 (EDMD1) [MIM:310300]. A degenerative myopathy characterized by weakness and atrophy of muscle without involvement of the nervous system, early contractures of the elbows Achilles tendons and spine, and cardiomyopathy associated with cardiac conduction defects.

  • Sequence similarities

    Contains 1 LEM domain.

  • Post-translational
    modifications

    Found in four different phosphorylated forms, three of which appear to be associated with the cell cycle.

  • Cellular localization

    Nucleus inner membrane. Nucleus outer membrane. Colocalized with BANF1 at the central region of the assembling nuclear rim, near spindle-attachment sites. The accumulation of different intermediates of prelamin-A/C (non-farnesylated or carboxymethylated farnesylated prelamin-A/C) in fibroblasts modify its localization in the nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • EDMD antibody
    • Emd antibody
    • EMD_HUMAN antibody
    • Emerin antibody
    • Emery Dreifuss muscular dystrophy antibody
    • STA antibody
    see all

Images

  • Western blot - Anti-Emerin antibody (ab40688)
    Western blot - Anti-Emerin antibody (ab40688)
    All lanes : Anti-Emerin antibody (ab40688) at 1 µg/ml

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : EMD knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 25, 29 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?



    Lanes 1-2: Merged signal (red and green). Green - ab40688 observed at 35 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab40688 Anti-Emerin antibody was shown to specifically react with Emerin in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266336 (knockout cell lysate ab257423) was used. Wild-type and Emerin knockout samples were subjected to SDS-PAGE. ab40688 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-Emerin antibody (ab40688)
    Western blot - Anti-Emerin antibody (ab40688)
    All lanes : Anti-Emerin antibody (ab40688) at 1 µg/ml

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : Emerin knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 25, 29 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab40688 observed at 35 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab40688 was shown to recognize Emerin in wild-type HAP1 cells as signal was lost at the expected MW in Emerin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Emerin knockout samples were subjected to SDS-PAGE. Ab40688 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 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-Emerin antibody (ab40688)
    Western blot - Anti-Emerin antibody (ab40688)
    All lanes : Anti-Emerin antibody (ab40688) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : Jurkat whole cell lysate (ab7899)
    Lane 4 : Jurkat nuclear extract lysate (ab14844)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 25, 29 kDa
    Observed band size: 33 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-Emerin antibody (ab40688)
    Immunocytochemistry/ Immunofluorescence - Anti-Emerin antibody (ab40688)
    ICC/IF image of ab40688 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab40688, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • Immunoprecipitation - Anti-Emerin antibody (ab40688)
    Immunoprecipitation - Anti-Emerin antibody (ab40688)
    Emerin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Emerin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab40688.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 33kDa; Emerin;non specific bands - 52 and 60kDa: We are unsure as to the identity of this extra band.

References (5)

Publishing research using ab40688? Please let us know so that we can cite the reference in this datasheet.

ab40688 has been referenced in 5 publications.

  • Essawy N  et al. An Emerin LEM-Domain Mutation Impairs Cell Response to Mechanical Stress. Cells 8:N/A (2019). PubMed: 31185657
  • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
  • Pawar S  et al. Efficient protein targeting to the inner nuclear membrane requires Atlastin-dependent maintenance of ER topology. Elife 6:N/A (2017). PubMed: 28826471
  • Ihalainen TO  et al. Differential basal-to-apical accessibility of lamin A/C epitopes in the nuclear lamina regulated by changes in cytoskeletal tension. Nat Mater 14:1252-1261 (2015). PubMed: 26301768
  • Kalendová A  et al. Nuclear actin filaments recruit cofilin and actin-related protein 3, and their formation is connected with a mitotic block. Histochem Cell Biol 142:139-52 (2014). PubMed: 25002125

Images

  • Western blot - Anti-Emerin antibody (ab40688)
    Western blot - Anti-Emerin antibody (ab40688)
    All lanes : Anti-Emerin antibody (ab40688) at 1 µg/ml

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : EMD knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 25, 29 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?



    Lanes 1-2: Merged signal (red and green). Green - ab40688 observed at 35 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab40688 Anti-Emerin antibody was shown to specifically react with Emerin in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266336 (knockout cell lysate ab257423) was used. Wild-type and Emerin knockout samples were subjected to SDS-PAGE. ab40688 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-Emerin antibody (ab40688)
    Western blot - Anti-Emerin antibody (ab40688)
    All lanes : Anti-Emerin antibody (ab40688) at 1 µg/ml

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : Emerin knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 25, 29 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab40688 observed at 35 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab40688 was shown to recognize Emerin in wild-type HAP1 cells as signal was lost at the expected MW in Emerin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Emerin knockout samples were subjected to SDS-PAGE. Ab40688 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 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-Emerin antibody (ab40688)
    Western blot - Anti-Emerin antibody (ab40688)
    All lanes : Anti-Emerin antibody (ab40688) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : Jurkat whole cell lysate (ab7899)
    Lane 4 : Jurkat nuclear extract lysate (ab14844)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 25, 29 kDa
    Observed band size: 33 kDa why is the actual band size different from the predicted?
  • Immunocytochemistry/ Immunofluorescence - Anti-Emerin antibody (ab40688)
    Immunocytochemistry/ Immunofluorescence - Anti-Emerin antibody (ab40688)
    ICC/IF image of ab40688 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab40688, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • Immunoprecipitation - Anti-Emerin antibody (ab40688)
    Immunoprecipitation - Anti-Emerin antibody (ab40688)
    Emerin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Emerin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab40688.
    Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
    Band: 33kDa; Emerin;non specific bands - 52 and 60kDa: We are unsure as to the identity of this extra band.

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