Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Musashi 1 / Msi1 antibody [EP1302] (ab52865)

Overview

  • Product name
    Anti-Musashi 1 / Msi1 antibody [EP1302]
    See all Musashi 1 / Msi1 primary antibodies
  • Description
    Rabbit monoclonal [EP1302] to Musashi 1 / Msi1
  • Host species
    Rabbit
  • Specificity
    Several customers have found that this antibody gives good results in mouse and rat however in our hands, we cannot obtain positive results. This antibody is therefore no longer covered by our Abpromise guarantee for use in mouse or rat.
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IHC-Fr, WB, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Chicken, Human, Quail
  • Immunogen

    Synthetic peptide within Human Musashi 1/ Msi1 (N terminal). The exact sequence is proprietary.
    (Peptide available as ab178003)

  • Positive control
    • PC12 cells, SH-SY-5Y cell lysate and human brain tissue.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52865 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
IHC-P 1/50 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/250 - 1/500.
IHC-Fr 1/100.
WB 1/1000. Detects a band of approximately 39 kDa (predicted molecular weight: 39 kDa).Can be blocked with Musashi 1 / Msi1 peptide (ab178003).

For unpurified use at 1/2000.

Flow Cyt 1/80.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/40.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      RNA binding protein that regulates the expression of target mRNAs at the translation level. Regulates expression of the NOTCH1 antagonist NUMB. Binds RNA containing the sequence 5'-GUUAGUUAGUUAGUU-3' and other sequences containing the pattern 5'-[GA]U(1-3)AGU-3'. May play a role in the proliferation and maintenance of stem cells in the central nervous system.
    • Tissue specificity
      Detected in fetal kidney, brain, liver and lung, and in adult brain and pancreas. Detected in hepatoma cell lines.
    • Sequence similarities
      Belongs to the Musashi family.
      Contains 2 RRM (RNA recognition motif) domains.
    • Domain
      The first RNA recognition motif binds more strongly to RNA compared to the second one.
    • Cellular localization
      Cytoplasm. Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • Msi 1 antibody
      • Msi1 antibody
      • MSI1H_HUMAN antibody
      • Musashi homolog 1 antibody
      • Musashi RNA binding protein 1 antibody
      • Musashi-1 antibody
      • Musashi1 antibody
      • RNA binding protein Musashi homolog 1 antibody
      • RNA-binding protein Musashi homolog 1 antibody
      see all

    Images

    • All lanes : Anti-Musashi 1 / Msi1 antibody [EP1302] (ab52865) at 1/2000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : MSI1 knockout HAP1 whole cell lysate
      Lane 3 : SH-SY5Y whole cell lysate

      Predicted band size: 39 kDa
      Observed band size: 39 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab52865 observed at 39 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab52865 was shown to specifically react with Musashi 1 / Msi1 in wild-type HAP1 cells as signal was lost in MSI1 knockout cells. Wild-type and MSI1 knockout samples were subjected to SDS-PAGE. Ab52865 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-Musashi 1 / Msi1 antibody [EP1302] (ab52865) at 1/1000 dilution (purified)

      Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
      Lane 2 : UMNSAH/DF-1 (chicken embryo fibroblast) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 39 kDa



      Blocking and diluting buffer: 5% NFDM/TBST.

    • Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Mushashi 1/ Msi1 with purified ab52865 at 1:80 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Immunocytochemistry/ Immunofluorescence analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Mushashi 1/ Msi1 with Purified ab52865 at 1:500 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200. Ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling Mushashi 1/ Msi1 with Purified ab52865 at 1:50 dilution (17.7 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.

    • Immunocytochemistry/Immunofluorescence analysis of Neuro-2a (mouse neuroblastoma) labelling Musashi 1 with purified ab52865 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised by 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    • Anti-Musashi 1 / Msi1 antibody [EP1302] (ab52865) at 1/2000 dilution (unpurified) + SH-SY-5Y cell lysate at 10 µg/ml

      Secondary
      goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 39 kDa
      Observed band size: 39 kDa

    • Immunohistochemistical detection (on formaldehyde/PFA-fixed paraffin-embedded sections) of Musashi 1 / Msi1 antibody [EP1302] (unpurified ab52865) on Quail Tissue sections (embryo d5/6 Brain stem T/S). Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Primary Antibody unpurified ab52865 incubated at 1/300 for 2 hours at RT. Secondary Antibody: Biotin labelled goat anti rabbit IgG (1/300).

    • Flow Cyt image of Musashi1 (ab52865)using Accutase digested single cell suspension of hESC. the cells were fixed adn permeabilized. The cells were incubated with unpurified ab52865 (1/20 using Prem/wash solution) for 30 mins at 23°C.

      See Abreview

    References

    This product has been referenced in:
    • Odle AK  et al. Association of Gnrhr mRNA With the Stem Cell Determinant Musashi: A Mechanism for Leptin-Mediated Modulation of GnRHR Expression. Endocrinology 159:883-894 (2018). Read more (PubMed: 29228137) »
    • Agoston AT  et al. Columnar-Lined Esophagus Develops via Wound Repair in a Surgical Model of Reflux Esophagitis. Cell Mol Gastroenterol Hepatol 6:389-404 (2018). Read more (PubMed: 30186929) »
    See all 27 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Q&A

    Answer

    Thank you very much for your interest our products.

    To our knowledge, neither ab21628 nor ab52865 have been tested in Xenopus. The homology of the immunogen of ab21628 with Xenopus laevis and tropicalis is 76%. The homology of the immunogen of ab52865 with Xenopus laevis and tropicalis is 80%. So actually, the homology with ab52865 is higher. We had incorrectly listed Xenopus as "predicted to react with" for ab21628, since 76% is quite low, so we thank you for bringing that to our attention.

    By participating in our AbTrial program you can now use our products in an untested application or species without financial risk.

    Simply follow these easy steps below to apply for our AbTrial Program:

    1. Reply to this email, letting us know you are interested in testing ab52865 in Xenopus.

    2. Our scientists will email you an inactive personal discount code for the value of the product.

    3. Purchase and test the product at the regular price.

    4. Submit your results, including your discount code in the additional notes section of your Abreview.

    5. Once the Abreview is submitted, the discount code will become active.

    6. Apply your discount code on your next order to receive that value off.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: https://www.abcam.com/abtrial.

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    Answer

    Thank you for your enquiry. I am afraid that using PC12 as a positive control information was not taken from any publication. The cell line was used in our laboratory testing and the results obtained were good. Therefore, we had that information stated on the datasheet as a positive control that customers can use. I did however manage to locate a publication on the subject on the internet, should you wish to refer. This publication reports Musashi expression in PC12 cells. Function and regulation of the mammalian Musashi mRNA translational regulator. Biochem Soc Trans. 2008 June; 36(Pt 3): 528–530. http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2562719 Our recommended incubation period and time for ICC/IF application is 1 hr room temperature, or overnight at 4 degrees. Please refer to our protocols page for more protocol details on the application. https://www.abcam.com/ps/pdf/protocols/icc.pdf I hope this information is helpful. If there is anything else that I can help you with, please do not hesitate to contact me.

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