Product nameAnti-EMSY antibody
See all EMSY primary antibodies
DescriptionRabbit polyclonal to EMSY
SpecificityThis antibody detects a band of the same size by Western blot as ab4579, which was raised with a recombinant fragment of EMSY with no sequence in common with the immunogen for this antibody (and was raised 7 years after this antibody). The band detected by this antibody also runs at the same height as full length in vitro translated EMSY. This confirms that the band detected is EMSY.
Tested applicationsSuitable for: ICC/IF, WB, IP, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Recombinant fragment derived from residues 100 - 200 of Human EMSY.
- MCF-7, MEF, 293T and U20S. The antibody also detects in vitro translated EMSY
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: 0.05% Sodium azide
Our Abpromise guarantee covers the use of ab123 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/3000. Detects a band of approximately 141 kDa (predicted molecular weight: 141 kDa). Variability in the size at which EMSY runs by Western blot has been experienced with different protein marker systems. EMSY has been observed to run at between 115 and 150kD. However, the independently raised ab4579 has been used to confirm the specificity of this antibody.|
|IHC-P||1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.|
- Information by UniProt
- C11orf30 antibody
- Chromosome 11 open reading frame 30 antibody
- Emsy antibody
WB using EMSY antibody (ab123) demonstrating the interaction between EMSY and BRCA2 by immunoprecipitation.
Endogenous coimmunoprecipitation of EMSY and BRCA2 from unmanipulated asynchronously dividing HeLa cells.
Immunoprecipitation with an unrelated antibody (anti-GFP), preimmune serum, ab123 or anti-BRCA2 antibody was followed by Western blotting with ab123.
Neither of the unrelated antibodies could coprecipitate EMSY, whereas an EMSY signal was detected in the immunoprecipitate obtained with the anti-BRCA2 antibody. The anti-EMSY antibody could also immunoprecipitate endogenous EMSY protein.
EMSY was immunoprecipitated using 0.5mg MCF7 whole cell extract, Rabbit polyclonal to EMSY diluted to 1/3,000 in RIPA buffer 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, MCF7 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 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab123.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 141kDa; EMSY
Image courtesy of Human Protein Atlas
ab123 staining EMSY in human fallopian tube tissue (nucleus of the glandular cells). Paraffin embedded tissue was incubated with ab123 (1/500 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab123 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
Upper panel: EMSY responds to DNA damage.
EMSY antibody (ab123) immunofluorescence staining of immortalized mouse wild-type embryonic fibroblasts before and after treatment with ionizing radiation (10 Gray at 1.8 Gy/minute, 250 kVp). EMSY assembles into nuclear speckles over several hours.
Lower panel: Costaining with a monoclonal mouse antibody to gamma-H2AX reveals that EMSY relocalizes to DNA damage sites.
Western blot of EMSY on MCF-7 cell lysate.
Lane 1: ab4579 at 1/500
Lane 2: ab123 at 1/500.
Variability in the size at which EMSY runs by Western blot has been experienced with different protein marker systems. EMSY has been observed to run at between 115 and 150kD (even though the predicted size is 141kD). However, that the same size band is detected by both ab123 and ab4579 (raised with immunogens of different sequence) confirms the specificity of these antibodies.
Secondary ab Lane 1: Rabbit polyclonal to Goat IgG H&L (HRP) ab6741 (1/5000)
Exposure time: 3 min.
Secondary ab Lane 2: Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed ab7090 (1/5000)
Exposure time: 10 sec.
Lysate at 20
Lane 1 - Exposure time : 3 min.
Lane 2 - Exposure time : 10 sec.
Variability in the size at which EMSY runs by Western blot has been exper
This product has been referenced in:
- Kolinjivadi AM et al. Smarcal1-Mediated Fork Reversal Triggers Mre11-Dependent Degradation of Nascent DNA in the Absence of Brca2 and Stable Rad51 Nucleofilaments. Mol Cell 67:867-881.e7 (2017). Read more (PubMed: 28757209) »
- Altinisik J et al. Expression of EMSY gene in sporadic ovarian cancer. Mol Biol Rep 38:359-63 (2011). WB ; Human . Read more (PubMed: 20349280) »