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  1. Link

    products/primary-antibodies/msi2-antibody-ep1305y-ab76148.pdf

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Epigenetics and Nuclear Signaling DNA / RNA Translation Regulation
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MSI2 antibody [EP1305Y] (ab76148)

  • Datasheet
  • SDS
Reviews (1)Q&A (2)References (34)

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Western blot - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Western blot - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Western blot - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Immunocytochemistry/ Immunofluorescence - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Flow Cytometry (Intracellular) - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Immunoprecipitation - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Western blot - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Immunocytochemistry/ Immunofluorescence - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Flow Cytometry (Intracellular) - Anti-MSI2 antibody [EP1305Y] (ab76148)
  • Anti-MSI2 antibody [EP1305Y] (ab76148)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1305Y] to MSI2
  • Suitable for: Flow Cyt (Intra), IP, ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free HRP

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Protein
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Recombinant Human MSI2 protein (ab167853)
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
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Anti-ANP32E antibody (ab5993)

View more associated products

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: Flow Cyt (Intra), IP, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Epitope

    Based on the immunogen sequence for this antibody, it is not predicted to detect the shorter isoforms of MSI2.
  • Positive control

    • WB: HeLa, A549 MCF7, HAP1, Rat brain, Human brain, A549 , SW480 and T47D cell lysates IHC-P: Human placenta, Human bladder carcinoma Tissue IP: T-47D cell lysate. ICC/IF: PC12 and MCF7 cells Flow Cyt (intra): T-47D and HeLa cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1305Y
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • Translation
    • Regulation
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Other
    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • DNA / RNA binding

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-MSI2 antibody [EP1305Y] (ab199067)
    • Alexa Fluor® 647 Anti-MSI2 antibody [EP1305Y] (ab199251)
    • HRP Anti-MSI2 antibody [EP1305Y] (ab199252)
    • Anti-MSI2 antibody [EP1305Y] - BSA and Azide free (ab221789)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human MSI2 protein (ab167853)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For unpurified use at 1/30

IP
1/50.
ICC/IF
1/100.
WB
1/1000 - 1/2000. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).
IHC-P (1)
1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

Notes
Flow Cyt (Intra)
1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For unpurified use at 1/30

IP
1/50.
ICC/IF
1/100.
WB
1/1000 - 1/2000. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

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.
  • Target information above from: UniProt accession Q96DH6 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 124540 Human
    • Entrez Gene: 76626 Mouse
    • Entrez Gene: 360596 Rat
    • Omim: 607897 Human
    • SwissProt: Q96DH6 Human
    • SwissProt: Q920Q6 Mouse
    • Unigene: 658922 Human
    • Unigene: 400451 Mouse
    • Unigene: 428433 Mouse
    • Unigene: 474737 Mouse
    see all
  • 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

  • Western blot - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Western blot - Anti-MSI2 antibody [EP1305Y] (ab76148)
    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.

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

    Lane 1 : Mouse brain lysates
    Lane 2 : Rat brain lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSI2 antibody [EP1305Y] (ab76148)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder carcinoma tissue sections labeling MSI2 with purified ab76148 at 1:500 dilution (2.14 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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

    Lane 1 : T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysates
    Lane 2 : A549 (Human lung carcinoma epithelial cell) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

  • Immunocytochemistry/ Immunofluorescence - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Immunocytochemistry/ Immunofluorescence - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling MSI2 with purified ab76148 at 1:100 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with None. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry (Intracellular) - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Flow Cytometry (Intracellular) - Anti-MSI2 antibody [EP1305Y] (ab76148)

    Intracellular Flow Cytometry analysis of T-47D (Human ductal breast epithelial tumor epithelial cell) cells labeling MSI2 with purified ab76148 at 1/100 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSI2 antibody [EP1305Y] (ab76148)

    Immunohistochemical analysis of paraffin-embedded human placenta with ab76148 at 1/100-1/250 dilution.

    Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Immunoprecipitation - Anti-MSI2 antibody [EP1305Y] (ab76148)
    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
  • Western blot - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Western blot - Anti-MSI2 antibody [EP1305Y] (ab76148)
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Immunocytochemistry/ Immunofluorescence - Anti-MSI2 antibody [EP1305Y] (ab76148)
    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.

  • Flow Cytometry (Intracellular) - Anti-MSI2 antibody [EP1305Y] (ab76148)
    Flow Cytometry (Intracellular) - Anti-MSI2 antibody [EP1305Y] (ab76148)

    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 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. 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.

     

  • Anti-MSI2 antibody [EP1305Y] (ab76148)
    Anti-MSI2 antibody [EP1305Y] (ab76148)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (34)

Publishing research using ab76148? Please let us know so that we can cite the reference in this datasheet.

ab76148 has been referenced in 34 publications.

  • Topchu I  et al. Musashi 2 (MSI2) expression as an independent prognostic biomarker in non-small cell lung cancer (NSCLC). J Thorac Dis 13:1370-1379 (2021). PubMed: 33841930
  • Makhov P  et al. Musashi-2 (MSI2) regulates epidermal growth factor receptor (EGFR) expression and response to EGFR inhibitors in EGFR-mutated non-small cell lung cancer (NSCLC). Oncogenesis 10:29 (2021). PubMed: 33723247
  • Montalbano M  et al. RNA-binding proteins Musashi and tau soluble aggregates initiate nuclear dysfunction. Nat Commun 11:4305 (2020). PubMed: 32855391
  • Wu X  et al. Targeting the interaction between RNA-binding protein HuR and FOXQ1 suppresses breast cancer invasion and metastasis. Commun Biol 3:193 (2020). PubMed: 32332873
  • Nguyen DTT  et al. HyperTRIBE uncovers increased MUSASHI-2 RNA binding activity and differential regulation in leukemic stem cells. Nat Commun 11:2026 (2020). PubMed: 32332729
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-MSI2 antibody [EP1305Y]

Excellent
Abreviews
Abreviews
abreview image
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
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 Aug 30 2017

Question

Blocking peptide for ab76148.
Thanks!

Read More

Abcam community

Verified customer

Asked on Jul 27 2012

Answer

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 hope this information is helpful to you.
Have a nice weekend and I wish you all the best with your research.

Read More

Abcam Scientific Support

Answered on Jul 27 2012

Question

Can you please provide some additional details about the immunogen for this MSI2 antibody? Will this antibody also detect the shorter forms of MSI2?

Read More

Abcam community

Verified customer

Asked on Jan 25 2012

Answer

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 know if you need any additional information.

Read More

Abcam Scientific Support

Answered on Jan 25 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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