Anti-Dicer 抗体 [13D6] - ChIP Grade (ab14601)

製品の概要

  • 製品名Anti-Dicer antibody [13D6] - ChIP Grade
    Dicer 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [13D6] to Dicer - ChIP Grade
  • 特異性We do not guarantee its use in WB with mouse lysates.
  • アプリケーション適用あり: ICC, IP, ICC/IF, IHC-P, WB, ChIP, Flow Cytmore details
  • 種交差性
    交差種: Human
    非交差種: Caenorhabditis elegans
  • 免疫原

    Synthetic peptide corresponding to Human Dicer.

  • ポジティブ・コントロール
    • WB: 10ng of recombinant dicer (using ECL or ECL Plus) in under one minute of exposure to film. ICC: Human Fibroblasts
  • 特記事項

    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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

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.

ターゲット情報

  • 機能Double-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.
  • 関連疾患Pleuropulmonary 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).
  • 配列類似性Belongs 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.
  • 細胞内局在Cytoplasm.
  • Information by UniProt
  • 参照データベース
  • 別名
    • DCR antibody
    • DCR1 antibody
    • Dicer 1 ribonuclease type III antibody
    • Dicer antibody
    • DICER_HUMAN antibody
    • DICER1 antibody
    • Double-strand-specific ribonuclease antibody
    • Endoribonuclease Dicer antibody
    • Helicase MOI antibody
    • Helicase with RNase motif antibody
    • HERNA antibody
    • KIAA0928 antibody
    • MNG1 antibody
    • RMSE2 antibody
    see all

Anti-Dicer antibody [13D6] - ChIP Grade 画像

  • 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 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) ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) ab216777 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • 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
    Goat anti-Mouse Green at 1/10000 dilution

    This 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.

  • 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.
  • 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 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.

  • All 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?)

    This image is courtesy of Jing Huang and Elliot Epner Oregon Health and Science University

Anti-Dicer antibody [13D6] - ChIP Grade (ab14601) 使用論文

This product has been referenced in:
  • Salzman DW  et al. miR-34 activity is modulated through 5'-end phosphorylation in response to DNA damage. Nat Commun 7:10954 (2016). WB ; Human . Read more (PubMed: 26996824) »
  • Dhir A  et al. Microprocessor mediates transcriptional termination of long noncoding RNA transcripts hosting microRNAs. Nat Struct Mol Biol 22:319-27 (2015). Human . Read more (PubMed: 25730776) »

See all 54 Publications for this product

Product Wall

Application Western blot
Sample Hamster Cell lysate - whole cell (VeroE6 (Kidney from African green monkey), BHK21 ()
Gel Running Conditions Reduced Denaturing (4-12% NuPage gel, MES buffer)
Loading amount 25000 cells
Specification VeroE6 (Kidney from African green monkey), BHK21 (
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 24°C
Username

Abcam user community

Verified customer

投稿 Jul 11 2016

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Sample Human Cell (Hela)
Specification Hela
Permeabilization Yes - 0.05% tritonx-100
Fixative Formaldehyde
Username

Dr. santosh chauhan

Verified customer

投稿 Apr 14 2014

The peptide immunogen was xxxxxxxx which corresponds to xxxxxxxx of the immature unprocessed sequence.

I would therefore suggest you use the following blocking peptide: Dicer peptide (ab24556)

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Mammary Tumor)
Gel Running Conditions Reduced Denaturing (8%)
Loading amount 60 µg
Specification Mammary Tumor
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 May 31 2013

Abreviews
Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (8%)
Sample Rat Tissue lysate - whole (Mammary Tumor)
Specification Mammary Tumor
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 May 30 2013

I am very pleased to hear you would like to test ab14601 in rat. This code will give you $379 off your next order before the expiration date. To redeem this offer, please submit an Abreview for rat and include this code in the “Additional Comment...

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Acording to UniProt, http://www.uniprot.org/uniprot/Q9UPY3, there are 2 isoforms of the Human Endoribonuclease Dicer (Gene name = Dicer1).
The isoform 2, also known as t-Dicer, differs from the canonical sequence as follows: 1789-1922: LRRSEED...

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Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from the last 2 vials of this antibody.

The details you have kindly provided will provide us with vital information for ou...

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Thank you for contacting Abcam.
The peptide used to generate this antibody was located between amino acids *****of the human form of the protein (SwissProt: Q9UPY3 Human).
Please let me know if there is anything else I can help you with, Free...

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Thank you very much for your call today and for letting us know about the trouble with this antibody.

Here areour suggestions for transferring high molecular weight proteins (>100 kDa):

For large proteins, transfer out of the gel...

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