Validated using a knockout cell line
Recombinant
RabMAb

Anti-MSI2 antibody [EP1305Y] (ab76148)

Overview

  • Product name
    Anti-MSI2 antibody [EP1305Y]
    See all MSI2 primary antibodies
  • Description
    Rabbit monoclonal [EP1305Y] to MSI2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide derived from Human MSI2 (UniProt: Q96DH6).

  • Epitope
    Based on the immunogen sequence for this antibody, it is not predicted to detect the shorter isoforms of MSI2.
  • Positive control
    • WB: HeLa, MCF7, HAP1, Rat brain, Human brain, SW480 and T47D cell lysates IHC-P: Human placenta tissue IP: T-47D cell lysate. ICC/IF: PC12 cells FC: HeLa cells.
  • 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 ab76148 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
IP 1/50.
ICC/IF Use a concentration of 1 µg/ml.
WB 1/1000 - 1/2000. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).Can be blocked with MSI2 peptide (ab136164).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Flow Cyt 1/30.

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

Target

  • Function
    RNA binding protein that regulates the expression of target mRNAs at the translation level. May play a role in the proliferation and maintenance of stem cells in the central nervous system.
  • Tissue specificity
    Ubiquitous; detected at low levels.
  • Involvement in disease
    Note=Chromosomal aberrations involving MSI2 may contribute to disease progression in chronic myeloid leukemia. Translocation t(7;17)(p15;q23) with HOXA9; translocation t(7;17)(q32-34;q23).
  • Sequence similarities
    Belongs to the Musashi family.
    Contains 2 RRM (RNA recognition motif) domains.
  • Post-translational
    modifications
    Phosphorylated.
  • Cellular localization
    Cytoplasm. Associated with polysomes.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ36569 antibody
    • MGC3245 antibody
    • Msi2 antibody
    • MSI2/HOXA9 fusion gene, included antibody
    • MSI2H antibody
    • MSI2H_HUMAN antibody
    • Musashi 2 antibody
    • Musashi homolog 2 antibody
    • Musashi RNA binding protein 2 antibody
    • Musashi, Drosophila, homolog of, 2 antibody
    • Musashi-2 antibody
    • RNA binding protein Musashi homolog 2 antibody
    • RNA-binding protein Musashi homolog 2 antibody
    • WD 40 repeat protein MSI2 antibody
    see all

Images

  • All lanes : Anti-MSI2 antibody [EP1305Y] (ab76148) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : MSI2 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : MCF7 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 35 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab76148 observed at 40 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab76148 was shown to specifically react with MSI2 in wild-type HAP1 cells as signal was lost in MSI2 knockout cells. Wild-type and MSI2 knockout samples were subjected to SDS-PAGE. Ab76148 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 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.

  • Immunohistochemical analysis of paraffin-embedded human placenta with ab76148 at 1/100-1/250 dilution.
  • All lanes : Anti-MSI2 antibody [EP1305Y] (ab76148) at 1/1000 dilution

    Lane 1 : T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate at 10 µg with 5% NFDM/TBST
    Lane 2 : T-47D whole cell lysate. ab76148 use as the capture antibody at 1:50 dilution. with 5% NFDM/TBST
    Lane 3 : T-47D whole cell lysate. ab172730 use as the capture antibody at 1:50 dilution. with 5% NFDM/TBST

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution (VeriBlot for IP secondary antibody (HRP))

    Observed band size: 35 kDa (why is the actual band size different from the predicted?)


    Exposure time: 5 seconds
  • All lanes : Anti-MSI2 antibody [EP1305Y] (ab76148) at 1/2000 dilution

    Lane 1 : Rat brain cell lysates
    Lane 2 : Human brain cell lysates
    Lane 3 : SW480 cell lysates
    Lane 4 : T47D cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 35 kDa
    Observed band size: 35 kDa

  • ICC/IF image of ab76148 stained PC12 cells. The cells were 4% formaldehyde 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 (ab76148, 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.

  • Overlay histogram showing HeLa cells stained with ab76148 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab76148, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in HeLa cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Liu Y  et al. Musashi-2 is a prognostic marker for the survival of patients with cervical cancer. Oncol Lett 15:5425-5432 (2018). Read more (PubMed: 29556294) »
  • Belew MS  et al. PLAG1 and USF2 Co-regulate Expression of Musashi-2 in Human Hematopoietic Stem and Progenitor Cells. Stem Cell Reports 10:1384-1397 (2018). WB ; Human . Read more (PubMed: 29641991) »

See all 12 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Pancreatic cancer tissues)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate buffer/trypsin
Permeabilization
No
Specification
Pancreatic cancer tissues
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 30 2017

Thank you for contacting us and for your interest in our products.
The blocking peptide for the anti-MSI2 antibody ab76148 is now available from our catalogue. The reference is ab136164 and the link to the datasheet is www.abcam.com/ab136164.
I...

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Thank you for contacting us.

The immunogen information for this antibody is considered proprietary. However, based on the sequence information, this antibody will not recognize the shorter isoforms of MSI2. I hope this helps, please let me kn...

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