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  1. Link

    drosha-antibody-ab12286.pdf

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Epigenetics and Nuclear Signaling DNA / RNA RNA Processing RNAi
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Anti-Drosha antibody (ab12286)

  • Datasheet
  • SDS
Reviews (9)Q&A (12)References (87)

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Immunocytochemistry - Anti-Drosha antibody - ChIP Grade (ab12286)
  • Western blot - Anti-Drosha antibody - ChIP Grade (ab12286)
  • Immunocytochemistry - Anti-Drosha antibody (ab12286)
  • Western blot - Anti-Drosha antibody - ChIP Grade (ab12286)
  • Immunocytochemistry - Anti-Drosha antibody (ab12286)
  • Immunocytochemistry - Anti-Drosha antibody (ab12286)

Key features and details

  • Rabbit polyclonal to Drosha
  • Suitable for: WB, ICC
  • Reacts with: Mouse, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Drosha antibody [EPR23046-123] (ab242147)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Drosha antibody
    See all Drosha primary antibodies
  • Description

    Rabbit polyclonal to Drosha
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICCmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human Drosha aa 1-100 (internal sequence).
    (Peptide available as ab12307)

  • General notes

    Three isoforms of 159, 156 and 143 kDa.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped 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 buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • RNAi
    • Epigenetics and Nuclear Signaling
    • RNAi
    • Dicer
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Nuclear Pore Complex

Associated products

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
WB (4)
Use a concentration of 1 µg/ml. Predicted molecular weight: 159 kDa.
ICC (1)
Use a concentration of 1 µg/ml.
Notes
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 159 kDa.
ICC
Use a concentration of 1 µg/ml.

Target

  • Function

    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.
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    Contains 1 DRBM (double-stranded RNA-binding) domain.
    Contains 2 RNase III domains.
  • Domain

    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.
  • Cellular localization

    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.
  • Target information above from: UniProt accession Q9NRR4 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 29102 Human
    • Entrez Gene: 14000 Mouse
    • Omim: 608828 Human
    • SwissProt: Q9NRR4 Human
    • SwissProt: Q5HZJ0 Mouse
    • Unigene: 97997 Human
    • Unigene: 293142 Mouse
    • Alternative names

      • 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

    Images

    • Immunocytochemistry - Anti-Drosha antibody - ChIP Grade (ab12286)
      Immunocytochemistry - Anti-Drosha antibody - ChIP Grade (ab12286)This image is courtesy of Ahmad M. Khalil and Daniel J. Driscoll, University of Florida College of Medicine Genetics Institute.
      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.
    • Western blot - Anti-Drosha antibody - ChIP Grade (ab12286)
      Western blot - Anti-Drosha antibody - ChIP Grade (ab12286)
      All lanes : Anti-Drosha antibody (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
      All lanes : 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?

    • Immunocytochemistry - Anti-Drosha antibody (ab12286)
      Immunocytochemistry - Anti-Drosha antibody (ab12286)

      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

    • Western blot - Anti-Drosha antibody - ChIP Grade (ab12286)
      Western blot - Anti-Drosha antibody - ChIP Grade (ab12286)
      All lanes : Anti-Drosha antibody (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
      All lanes : 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.
    • Immunocytochemistry - Anti-Drosha antibody (ab12286)
      Immunocytochemistry - Anti-Drosha antibody (ab12286)

      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.

    • Immunocytochemistry - Anti-Drosha antibody (ab12286)
      Immunocytochemistry - Anti-Drosha antibody (ab12286)

      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.

    Protocols

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (87)

    Publishing research using ab12286? Please let us know so that we can cite the reference in this datasheet.

    ab12286 has been referenced in 87 publications.

    • Zhu M  et al. Novel roles of an intragenic G-quadruplex in controlling microRNA expression and cardiac function. Nucleic Acids Res 49:2522-2536 (2021). PubMed: 33561291
    • Fóthi Á  et al. Tissue-specific and transcription-dependent mechanisms regulate primary microRNA processing efficiency of the human chromosome 19 MicroRNA cluster. RNA Biol N/A:1-11 (2020). PubMed: 33052778
    • Zhang L  et al. RNA helicase p68 inhibits the transcription and post-transcription of Pkd1 in ADPKD. Theranostics 10:8281-8297 (2020). PubMed: 32724471
    • Baldini E  et al. Dysregulation of microRNA expression in diabetic skin. J Dermatol Sci 98:186-194 (2020). PubMed: 32402513
    • Milon A  et al. Effect of estrogen-related receptor silencing on miRNA protein machinery expression, global methylation, and deacetylation in bank vole (Myodes glareolus) and mouse tumor Leydig cells. Theriogenology 139:178-190 (2019). PubMed: 31421412
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 21 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Drosha antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Ovary and Fallopian Tube and Uterine)
    Antigen retrieval step
    Heat mediated
    Permeabilization
    No
    Specification
    Ovary and Fallopian Tube and Uterine
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Eunyoung Park

    Verified customer

    Submitted Dec 10 2020

    Immunoprecipitation abreview for Anti-Drosha antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Immuno-precipitation step
    Protein G
    Sample
    Human Cell lysate - nuclear (A549)
    Specification
    A549
    Total protein in input
    25 µg
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 20 2015

    Western blot

    Average
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted May 27 2013

    Western blot

    Average
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted May 21 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Drosha antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Kai Orton

    Verified customer

    Submitted May 28 2012

    Question

    I performed a western blot with this antibody twice with two different conditions. Both were using protein isolated from three mouse embryonic cell lines (two un-differentiated, one derived into motor neurons). 25ug/lane were loaded for the undifferentiated mESCs and 40ug for the mESC-dervied motor neuron samples (for both westerns). First western: Blocked with 5% milk 30min. Added anti-Drosha antibody 1:1000 (3ug/3ml) in 5% milk. incubated o/n at 4degC on shaker. Washed 3x with PBS+0.5%tween. Added secondary antibody (anti-Rabbit IgG HRP conjugated from donkey, other company) at 1:6000 in 5%milk. Incubated at RT for 1 hour. Washed 3x with PBS+0.5%tween, 2x in PBS. Developed using ECL method. Even after 10 minutes of developing, I could not see any bands whatsoever.

    Second western: Blocked with 0.5%milk 30min. Added anti-Drosha antibody 1:500 (6ug/3ml) in PBS. incubated 4 hours RT on shaker. Washed 3x with PBS. Added secondary antibody (anti-Rabbit IgG HRP conjugated from donkey, from other company) at 1:6000 in PBS. Incubated at RT for 1 hour. Washed 3x with PBS. Developed using ECL method. Saw multiple bands at 62kDa, 100kDa, doublet at >250kDa in the two un-differentiated mouse ESC lines. In the differentiated line, I saw bands at 90kDa, 150kDa, 200kDa. Some other faint bands were observed in the lanes. However, there was no band at 150 or 170kDa (the predicted and observed sizes of Drosha) across all three of my samples.
    These results suggest that I am not detecting Drosha. I would expect a band at 150kDa (or 170kDa) across all of my protein samples (all are isolated from mouse cells). However, there are many strong bands at other sizes. I would like a refund/credit, since this antibody was recommended for use on mouse and I do not have consistent results in mouse.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 14 2013

    Answer


    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.

    Read More

    Abcam Scientific Support

    Answered on Jan 14 2013

    Question

    Thank you , I appreciate your willingness to replace the antibody.
    I would like to request the PROX1 antibody cat# 38692 as a replacement. Please contact me if you require more information.
    Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on Aug 02 2012

    Answer

    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 antibody, or if there is anything else I can help you with.
    I hope you are having a great day and also that your experiments co-operate.

    Read More

    Abcam Scientific Support

    Answered on Aug 02 2012

    Question

    Our experience with the antibody:
    We used this antibody for western blot analysis to evaluate cell lines that differ in Drosha expression using qPCR. We also used MCF-7 lysates as a positive control. All samples were lysed in either RIPA or EB. We loaded up to 120 mcg per lane and never observed a band. The secondary antibody works fine with several other primary antibodies. We made several attempts and had more than one person test the antibody but never got it to work.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 02 2012

    Answer

    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 either send a replacement. Please let me know the catalogue number of the antibody you would like to receive as a replacement and I will have it sent out as soon as I can.
    I look forward to your reply and helping resolve this issue.

    Read More

    Abcam Scientific Support

    Answered on Aug 02 2012

    Question

    Please see customers inquiry regarding Anti-Drosha antibody - ChIP Grade (ab12286) below:



    I was wondering whether the following antibody cross-reacts with Drosophila Drosha. It is not in the description, but you never know.



    Looking forward for your response.





    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Jun 05 2012

    Answer

    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 experiments with drosophila.
    Unfortunately, the other two anti Drosha antibodies (ab58589 and ab85027) we have in our catalog at the moment also do not show any promise of a cross reaction with drosophila.
    The homology is lower than 45% in all cases.
    I am sorry I did not have better news on this occasion and hope this information is nevertheless helpful for your customer.

    Read More

    Abcam Scientific Support

    Answered on Jun 05 2012

    Question

    Thank you for sending us a new Drosha antibody. We had a chance to try it out, I have attached our western blot results.
    As you will see, we were not able to have a band around 179-190 as per protein molecular weight.
    Our fist two lanes are Nuclear extracts and the last two are Cytoplasmic ones. Drosha is only supposed to be in the nucleus, but our cytoplasmic samples are showing plenty of other bands.
    What do you think could be happening here? Is there a cleavage that happens and this antibody is showing it around 75KDa? Could we use this band as our reference?
    On another related note, we tried the same antibody for immuno-florescence and it worked very beautifully!!! It clearly was staining only on the nucleus.
    So this is really strange, as Abcam suggests that this Ab is suitable for both WB and IF, but WB result are really not happening....
    Please advice.

    Read More

    Abcam community

    Verified customer

    Asked on May 04 2012

    Answer

    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 in WB as ab58589. Would you like to give that one a try? Or we can refund your original purchase of ab12286.

    https://www.abcam.com/Drosha-antibody-ab58589.html

    Please let me know which you would prefer. I look forward to your reply.

    Read More

    Abcam Scientific Support

    Answered on May 04 2012

    1-10 of 21 Abreviews or Q&A

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