製品の概要

  • 製品名Anti-Drosha antibody - ChIP Grade
    Drosha 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Drosha - ChIP Grade
  • アプリケーション適用あり: ICC/IF, IP, WB, ICC, IHC-P, ChIPmore details
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    Synthetic peptide derived from within residues 1 - 100 of Human Drosha.

    (Peptide available as ab12307.)

  • ポジティブ・コントロール
    • HeLa cells
  • 特記事項Three isoforms of 159, 156 and 143 kDa.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab12286 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
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 159 kDa.
ICC Use at an assay dependent concentration.
IHC-P 1/100.
ChIP Use at an assay dependent concentration.

ターゲット情報

  • 機能Ribonuclease III double-stranded (ds) RNA-specific endoribonuclease that is involved in the initial step of microRNA (miRNA) biogenesis. Component of the microprocessor complex that is required to process primary miRNA transcripts (pri-miRNAs) to release precursor miRNA (pre-miRNA) in the nucleus. Within the microprocessor complex, DROSHA cleaves the 3' and 5' strands of a stem-loop in pri-miRNAs (processing center 11 bp from the dsRNA-ssRNA junction) to release hairpin-shaped pre-miRNAs that are subsequently cut by the cytoplasmic DICER to generate mature miRNAs. Involved also in pre-rRNA processing. Cleaves double-strand RNA and does not cleave single-strand RNA. Involved in the formation of GW bodies.
  • 組織特異性Ubiquitous.
  • 配列類似性Contains 1 DRBM (double-stranded RNA-binding) domain.
    Contains 2 RNase III domains.
  • ドメインThe 2 RNase III domains form an intramolecular dimer where the domain 1 cuts the 3'strand while the domain 2 cleaves the 5'strand of pri-miRNAs, independently of each other.
  • 細胞内局在Nucleus. Nucleus > nucleolus. A fraction is translocated to the nucleolus during the S phase of the cell cycle. Localized in GW bodies (GWBs), also known as P-bodies.
  • Information by UniProt
  • 参照データベース
  • 別名
    • DROSHA antibody
    • Drosha double stranded RNA specific endoribonuclease antibody
    • Drosha ribonuclease type III antibody
    • Etohi2 antibody
    • HSA242976 antibody
    • Nuclear RNase III Drosha antibody
    • p241 antibody
    • Protein Drosha antibody
    • Putative protein p241 which interacts with transcription factor Sp1 antibody
    • Putative ribonuclease III antibody
    • RANSE3L antibody
    • Ribonuclease 3 antibody
    • Ribonuclease III antibody
    • Ribonuclease III nuclear antibody
    • Ribonuclease type III nuclear antibody
    • RibonucleaseIII antibody
    • RN 3 antibody
    • RN3 antibody
    • RNase 3 antibody
    • RNase III antibody
    • RNase3 antibody
    • RNASE3L antibody
    • RNaseIII antibody
    • RNASEN antibody
    • RNC_HUMAN antibody
    see all

Anti-Drosha antibody - ChIP Grade 画像

  • Human female amniocytes immunostained with ab12286 Drosha (FITC) (1/25 dilution). The DNA is labelled red with propidium iodide. This image was submitted as part of a review by Ahmad Khalil.
  • ab12286 at a 1/25 dilution staining human HeLa cells by immunocytochemistry. The antibody was incubated with the cells for 1 hour and then detected using a Cy3 conjugated donkey anti-rabbit polyclonal antibody.

    This image is courtesy of an Abreview submitted on 7 February 2006.

    See Abreview

  • All lanes : Anti-Drosha antibody - ChIP Grade (ab12286) 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

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/15000 dilution

    Performed under reducing conditions.

    Predicted band size : 159 kDa
    Observed band size : 170 kDa (why is the actual band size different from the predicted?)
  • All lanes : Anti-Drosha antibody - ChIP Grade (ab12286) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Nuclear Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 159 kDa
    Observed band size : 170 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 115 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
    Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
  • ICC/IF image of ab12286 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab12286, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • ICC/IF image of ab12286 stained Hela cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab12886, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Anti-Drosha antibody - ChIP Grade (ab12286) 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) »
  • Jiang FZ  et al. Mutant p53 induces EZH2 expression and promotes epithelial-mesenchymal transition by disrupting p68-Drosha complex assembly and attenuating miR-26a processing. Oncotarget 6:44660-74 (2015). IP . Read more (PubMed: 26587974) »

See all 36 Publications for this product

Product Wall

Application Immunoprecipitation
Immuno-precipitation step Protein G
Sample Human Cell lysate - nuclear (A549)
Specification A549
Total protein in input 25 µg
Username

Abcam user community

Verified customer

投稿 Feb 20 2015

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing (13.5%)
Sample Mouse Cell lysate - whole cell (Mouse N2A neuroblastoma)
Specification Mouse N2A neuroblastoma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 May 27 2013

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (6%)
Sample Human Cell lysate - whole cell (human osteosarcoma cell line U2OS)
Specification human osteosarcoma cell line U2OS
Blocking step Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 May 21 2013


I can offer you either a free of charge replacement with another Drosha-antibody (e.g. ab135956) or an antibody to a different target. Alternatively, a credit or refund can be arranged.

Thank you for your reply.
As requested, I am sending you ab38692 as a free of charge replacement for ab12286 that did not work for you in western blotting. Your new order number is *.
Please let me know if you experience any issues with the new...

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Thank you for your reply.
The antibody, ab12286 is covered under our Abpromise for six months and is guaranteed to work in western blot on rabbit samples . Since we cannot resolve this issue you are having with the antibody then I would be happy to...

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Thank you and your customer for your inquiry.
I checked the homology between the immunogen sequence of ab12286 and drosophila and unfortunately have to confirm that there is no significant homology.
Therefore I cannot recommend ab12286 for ex...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Mouse Brain Day 25)
Specification Mouse Brain Day 25
Fixative neutral buffered formalin
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Permeabilization Yes - TBS-Tween (0.1%)
Blocking step Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Username

Dr. Kai Orton

Verified customer

投稿 May 28 2012

Thank you for your reply.

I am sorry to hear that the replacement antibody didn't work well in WB. I'm not sure why the antibody is showing so many non-specific bands.

We have a different anti-drosha antibody tested in human samples...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"