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Immunofluorescence - EPC1 antibody (ab5514)
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- EPC1 antibody (ab5514)
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Western blot - EPC1 antibody (ab5514)
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Product Name
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EPC1 antibody
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to EPC1
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 550 - 650 of Human EPC1.
(Peptide available as ab24396.)
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Reacts with
(species key)
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Hu
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Tested applications
(see key)
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ICC/IF, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC: 1/50.
IF: 1/50.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Nuclear
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Research areas
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Chromatin and Nuclear Signaling >> Chromatin Modifying Enzymes >> Acetylation >> HAT
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Relevance
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EPC1 is a component of the NuA4 histone acetyltransferase (HAT) complex which is involved in transcriptional activation of select genes, principally by acetylation of nucleosomal histone H4 and H2A. This modification may both alter nucleosome-DNA interactions and promote interaction of the modified histones with other proteins which positively regulate transcription. This complex may be required for the activation of transcriptional programs associated with oncogene and proto-oncogene mediated growth induction, tumor suppressor mediated growth arrest and replicative senescence, apoptosis, and DNA repair. NuA4 may also play a direct role in DNA repair when directly recruited to sites of DNA damage.
EPC1 is a component of the NuA4 histone acetyltransferase complex which contains the catalytic subunit HTATIP/TIP60 and the subunits EP400, TRRAP/PAF400, BRD8/SMAP, EPC1, DMAP1/DNMAP1, RUVBL1/TIP49, RUVBL2, ING3, actin, ACTL6A/BAF53A, MORF4L1/MRG15, MORF4L2/MRGX, MRGBP and YEATS4/GAS41. HTATTIP/TIP60, EPC1, and ING3 together constitute a minimal HAT complex termed Piccolo NuA4. The NuA4 complex also interacts with MYC and the adenovirus E1A protein. EPC1 also interacts with RFP1/TRIM27.
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Database links
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The links below go to external sites and will open in a new browser window Entrez Gene | | 80314 (Human)
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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0.500 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Notes
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From initial efforts, this antibody does not appear to work convincingly in Western Blot. Collaborative data has also found that this ab does not work in WB even with an overexpressed flagged form of EPC1.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this EPC1 antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for EPC1 antibody images, references, products related to ab5514 and other tools.
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EPC1 antibody images:
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 Immunofluorescence - EPC1 antibody (ab5514)
Specificity of ab5514 against EPC1
A and B: Detection using indirest immunofluorescence of the signal corresponding to EPC1 in EPC1 in 293 Cells were fixed using 4% formaldehyde blocked with PBS containing 3% milk and 0.5% Triton x-100 (sigma) incubated for 1 hour at 37 C with ab5514 at a 1/50 dilution. The cells were then washed three times and incubated for 1 hour at 37 C with a donkey anti-rabbit antibody coupled with Alexa Flour 488 (Molecular probes) at a 1/500 dilution, washed three times, stained with DAPI and mounted with Vectasheild(Abcyss). A: Acquisition on a CCD camera B: Confocal microscopy (3um stack along Z axis).
C. Detection using indirect immunofluoresence of the signals corresponding to the staining with the anti-FLAG mouse antibody (left, red) and the Abcam ab5514 directed against EPC1 (middle, green). 29GT cells were fixed 24 hours after their transfection with a vector containing a FLAG form of EPC1 sequence, using Lipofectamine 2000 transfection procedure (Invitrogen). Mouse anti-FLAG antibody was used at a 1/500 dilution and was detected with donkey anti-mouse antibody coupled with Alexa Flour 488 (Molecular probes) at a 1/500 dilution. Ab5514 was detected using a donkey anti-rabbit antibody coupled with Alexa Flour 488 (Molecular probes) at a 1/500 dilution. Confocal microscopy.
The endogenous EPC1 protein looks both cytoplasmic and nuclear in 293T cells, which is suprising. However, the few cells that survive the overexpression of a flagged form of the protein display an essentially nuclear Flag signal.
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- EPC1 antibody (ab5514)
Verification of the specificity of ab5514 against EPC1 The siRNA that extinguishes EPC1 antibody-associated signal were independently validated using quantitative RT-PCR.
D: Detection using indirect immunofluorescence of the signal corresponding to EPC1 in 293 cells. Cells were transfected with a control shRNA directed against luciferase (Siluc, left) or two siRNAs directed againstEPC1 (siR1EPC1, middle, and siR2EPC1, right). 24 hours after transfection cells were fixed, blocked, incubated for 1 hr at 37 C with ab5514 at a 1/50 dilution. The cells were then washed three times and incubated for 1 hour at 37 C with a donkey anti-rabbit antibody coupled with Alexa Flour 488 (Molecular probes) at a 1/500 dilution, washed three times, stained with DAPI and mounted with Vectasheild(Abcyss). E: Efficiency of expression knock-down by the SiRNAs against EPC1. Transfection was down as described in D. Quantification of EPC1 mRNA level, expressed as a ratio to RPL19 mRNA and compared to the control (siRI Luc transfection), using quantitative RT-PCR. Genevive Fourel
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Western blot - EPC1 antibody (ab5514)
All lanes : EPC1 antibody (ab5514) at 2 µg/ml
Lane 1 : Whole cell lysate from HEK293 cell line
Lane 2 : Whole cell lysate from HeLa cell line
Lane 3 : Whole cell lysate from U20S cell line
Lysates/proteins at 30 µg per lane.
Secondary
HRP-conjugated pig anti rabbit Ig
Performed under reducing conditions.
Exposure time : 15 minutes
Detection method: Supersignal West Dura.
Note: There are 3 characterised isoforms of EPC1 which give rise to proteins of 93 kDa, 90 kDa and 85 kDa (Swissprot). This antibody detects a species of ~85 kDa in all lysates, including HEK293 cells which are known to express EPC1. The clean blots, together with the IF data (see datasheet), suggests that this antibody is specific to EPC1 and will work in western blotting. This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.
See Abreview
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References for EPC1 antibody (ab5514)
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Stankunas K, Berger J, Ruse C, Sinclair DA, Randazzo F, Brock HW. The enhancer of polycomb gene of Drosophila encodes a chromatin protein conserved in yeast and mammals. Development. 1998 Oct;125(20):4055-66.
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Search PubMed (MEDLINE) for references to EPC1
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EPC1 antibody - more information
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