Anti-Emerin antibody (ab40725)
Key features and details
- Rabbit polyclonal to Emerin
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Emerin antibody
See all Emerin primary antibodies -
Description
Rabbit polyclonal to Emerin -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB Human -
Immunogen
Synthetic peptide corresponding to Human Emerin aa 200 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available asab40724) -
Positive control
- ab40725 gave a positive result in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK293 (Human embryonic kidney cell line) ab40725 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)
Properties
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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 concentrationConcentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40725 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB HumanAll applications MouseRatCowApplication Abreviews Notes 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).Notes 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
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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
modificationsFound 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
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Database links
- Entrez Gene: 399681 Cow
- Entrez Gene: 2010 Human
- Entrez Gene: 13726 Mouse
- Entrez Gene: 25437 Rat
- Omim: 300384 Human
- SwissProt: P50402 Human
- SwissProt: O08579 Mouse
- SwissProt: Q63190 Rat
see all -
Alternative names
- EDMD antibody
- Emd antibody
- EMD_HUMAN antibody
see all
Images
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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 - ab40725 observed at 35 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab40725 was shown to specifically react with Emerin in wild-type HAP1 cells as signal was lost in Emerin knockout cells. Wild-type and Emerin knockout samples were subjected to SDS-PAGE. Ab40725 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.
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All lanes : Anti-Emerin antibody (ab40725) 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)
Lane 5 : Hep G2 whole cell lysate (ab7900)
Lane 6 :Hep G2 nuclear extract lysate (ab14660)
Lane 7 :A-431 whole cell lysate (ab7909)
Lane 8 :HEK-293 whole cell lysate (ab7902)
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: 35 kDa why is the actual band size different from the predicted?
Additional bands at: 46 kDa. We are unsure as to the identity of these extra bands. -
ICC/IF image of ab40725 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab40725, 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).
Protocols
Datasheets and documents
References (1)
ab40725 has been referenced in 1 publication.
- Khanna R et al. E3 ubiquitin ligase RNF123 targets lamin B1 and lamin-binding proteins. FEBS J 285:2243-2262 (2018). PubMed: 29676528
Images
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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 - ab40725 observed at 35 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab40725 was shown to specifically react with Emerin in wild-type HAP1 cells as signal was lost in Emerin knockout cells. Wild-type and Emerin knockout samples were subjected to SDS-PAGE. Ab40725 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.
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All lanes : Anti-Emerin antibody (ab40725) 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)
Lane 5 : Hep G2 whole cell lysate (ab7900)
Lane 6 :Hep G2 nuclear extract lysate (ab14660)
Lane 7 :A-431 whole cell lysate (ab7909)
Lane 8 :HEK-293 whole cell lysate (ab7902)
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: 35 kDa why is the actual band size different from the predicted?
Additional bands at: 46 kDa. We are unsure as to the identity of these extra bands. -
ICC/IF image of ab40725 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab40725, 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).