Product nameAnti-Emerin antibody
See all Emerin primary antibodies
DescriptionRabbit polyclonal to Emerin
Tested applicationsSuitable for: IP, ICC/IF, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat, Cow
- ab40688 gave a positive result in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) HepG2 (Human hepatocellular liver carcinoma cell line) A431 (Human epithelial carcinoma cell line) ab40688 gave a positive result in the following nuclear lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) HepG2 (Human hepatocellular liver carcinoma cell line)
Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
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.
|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).|
FunctionStabilizes 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 specificitySkeletal muscle, heart, colon, testis, ovary and pancreas.
Involvement in diseaseDefects 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 similaritiesContains 1 LEM domain.
modificationsFound in four different phosphorylated forms, three of which appear to be associated with the cell cycle.
Cellular localizationNucleus 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
- EDMD antibody
- Emd antibody
- EMD_HUMAN antibody
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Emerin knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
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.
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.
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?
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).
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.
This product has been referenced in:
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). Read more (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). Read more (PubMed: 28826471) »