Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)
- Datasheet
- Specific References (99)
- Protocols
Overview
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Product nameAnti-Dicer antibody [13D6] - ChIP Grade
See all Dicer primary antibodies -
DescriptionMouse monoclonal [13D6] to Dicer - ChIP Grade
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Host speciesMouse
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SpecificityWe do not guarantee its use in WB with mouse lysates.
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Tested applicationsSuitable for: ICC, IP, ICC/IF, IHC-P, WB, ChIP, Flow Cytmore details
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Species reactivityReacts with: Human
Does not react with: Caenorhabditis elegans -
Immunogen
Synthetic peptide corresponding to Human Dicer.
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Positive control
- WB: 10ng of recombinant dicer (using ECL or ECL Plus) in under one minute of exposure to film. ICC: Human Fibroblasts
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General notes
To our knowledge the localization of Dicer remains to be fully determined and it appears that its localization can be nuclear and/or cytoplasmic. Please see reference Meltzer, 2005 for a reference to a cytoplasmic localisation (figure 1 of paper), as supported with the staining seen in the image on this datasheet.
Please note this antibody gave negative results in WB with mouse fibroblast cell line lysates. We do not guarantee its use in WB with mouse lysates.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading... -
PurityAmmonium Sulphate Precipitation
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ClonalityMonoclonal
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Clone number13D6
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IsotypeIgG2a
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Research areas
Associated products
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry kits
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab14601 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 |
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| EMSA | Use at an assay dependent concentration. PubMed: 20972213 | |
| ICC | 1/20 - 1/100. | |
| IP | Use at an assay dependent concentration. PubMed: 24668803 | |
| ICC/IF | Use at an assay dependent concentration. | |
| IHC-P | Use at an assay dependent concentration. PubMed: 18327211 | |
| WB | 1/100 - 1/2000. Please note this antibody gave negative results in WB with mouse fibroblast cell line lysates. We do not guarantee its use in WB with mouse lysates. |
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| ChIP | Use at an assay dependent concentration. | |
| Flow Cyt | Use 0.1µg for 106 cells. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Target
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FunctionDouble-stranded RNA (dsRNA) endoribonuclease playing a central role in short dsRNA-mediated post-transcriptional gene silencing. Cleaves naturally occurring long dsRNAs and short hairpin pre-microRNAs (miRNA) into fragments of twenty-one to twenty-three nucleotides with 3' overhang of two nucleotides, producing respectively short interfering RNAs (siRNA) and mature microRNAs. SiRNAs and miRNAs serve as guide to direct the RNA-induced silencing complex (RISC) to complementary RNAs to degrade them or prevent their translation. Gene silencing mediated by siRNAs, also called RNA interference, controls the elimination of transcripts from mobile and repetitive DNA elements of the genome but also the degradation of exogenous RNA of viral origin for instance. The miRNA pathway on the other side is a mean to specifically regulate the expression of target genes.
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Involvement in diseasePleuropulmonary blastoma
Goiter multinodular 1, with or without Sertoli-Leydig cell tumors
Rhabdomyosarcoma, embryonal, 2
DICER1 mutations have been found in uterine cervix embryonal rhabdomyosarcoma, primitive neuroectodermal tumor, Wilms tumor, pulmonary sequestration and juvenile intestinal polyp (PubMed:21882293). Somatic missense mutations affecting the RNase IIIb domain of DICER1 are common in non-epithelial ovarian tumors. These mutations do not abolish DICER1 function but alter it in specific cell types, a novel mechanism through which perturbation of microRNA processing may be oncogenic (PubMed:22187960). -
Sequence similaritiesBelongs to the helicase family. Dicer subfamily.
Contains 1 Dicer dsRNA-binding fold domain.
Contains 1 DRBM (double-stranded RNA-binding) domain.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain.
Contains 1 PAZ domain.
Contains 2 RNase III domains. -
Cellular localizationCytoplasm.
- Information by UniProt
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Database links
- Entrez Gene: 23405 Human
- Omim: 606241 Human
- SwissProt: Q9UPY3 Human
- Unigene: 738957 Human
- Unigene: 87889 Human
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Alternative names
- DCR antibody
- DCR1 antibody
- Dicer 1 ribonuclease III antibody
see all
Images
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![Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)](https://www.abcam.com/ps/products/14/ab14601/Images/ab14601-256152-ab14601KOWB.jpg)
Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Dicer knockout HAP1 cell lysate (20 µg)Lanes 1 and 2: Merged signal (red and green). Green - ab14601 observed at 240 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab14601 was shown to specifically react with Dicer in wild-type HAP1 cells. No band was observed when Dicer knockout samples were used. Wild-type and Dicer knockout samples were subjected to SDS-PAGE. ab14601 and ab181602 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging. -
![Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)](https://www.abcam.com/ps/products/14/ab14601/Images/ab14601-255505-ab14601GR185750812000.jpg)
Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)All lanes : Anti-Dicer antibody [13D6] - ChIP Grade (ab14601) at 1/2000 dilution (incubated overnight at 4°C.)
Lane 1 : NIH 3T3
Lane 2 : A549
Lane 3 : HepG2
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Mouse Green at 1/10000 dilutionThis blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with ab14601 overnight at 4°C. Antibody binding was detected using IR-labelled goat anti-mouse Ab at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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![Flow Cytometry - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)](https://www.abcam.com/ps/products/14/ab14601/Images/ab14601-7-ab14601FC.jpg)
Flow Cytometry - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)Overlay histogram showing HEK293 cells stained with ab14601 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab14601, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. -
![Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)](https://www.abcam.com/ps/products/14/ab14601/Images/ab14601-238523-Figure-1.jpg)
Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)HepG2 cell lysate at 20 µg
Developed using the ECL technique.
Performed under reducing conditions.
Additional bands at: 100 kDa (possible non-specific binding), 60 kDa (possible non-specific binding) -
![ChIP - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)](https://www.abcam.com/ps/products/14/ab14601/Images/ab14601_5.jpg)
ChIP - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)This image is courtesy of Elliot Epner MD PhD, Oregon Health and Science University, PortlandChIP analysis of Dicer protein binding at chicken ß-globin regulatory elements. Chromatin from human erythroleukemia (K562) cells containing chicken chromosomes with normal (FRK2) or mutant (HS1-K) ß-globin loci and chicken DT40 cells was immunprecipitated with antibodies to Dicer (ab14601). PCR analysis of immunoprecipitated chromatin was carried out using primers to the chicken ß-globin regulatory elements. Mouse IgG served as control. 10% of input DNA was utilized.
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![Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)](https://www.abcam.com/ps/products/14/ab14601/Images/ab14601_7.jpg)
Western blot - Anti-Dicer antibody [13D6] - ChIP Grade (ab14601)This image is courtesy of Jing Huang and Elliot Epner Oregon Health and Science UniversityAll lanes : Anti-Dicer antibody [13D6] - ChIP Grade (ab14601) at 2.5 µg/ml
Lane 1 : human B cell lymphoma lysate
Lane 2 : human B cell lymphoma lysate infected with control shRNA retrovirus
Lane 3 : human B cell lymphoma lysate infected with Dicer knockdown shRNA virus
Lysates/proteins at 2.5 µg/ml per lane.
Observed band size: 225 kDa (why is the actual band size different from the predicted?)
Protocols
References
This product has been referenced in:
- Yang Z et al. Dicer reprograms stromal fibroblasts to a pro-inflammatory and tumor-promoting phenotype in ovarian cancer. Cancer Lett 415:20-29 (2018). WB, IF . Read more (PubMed: 29199004) »
- Tarallo R et al. The nuclear receptor ERß engages AGO2 in regulation of gene transcription, RNA splicing and RISC loading. Genome Biol 18:189 (2017). WB ; Human . Read more (PubMed: 29017520) »
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