Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1305Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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 |
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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 |
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IP |
1/50.
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ICC/IF |
1/100.
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WB |
1/1000 - 1/2000. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).
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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 |
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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
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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
modificationsPhosphorylated. -
Cellular localization
Cytoplasm. Associated with polysomes. - Information by UniProt
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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
see all -
Alternative names
- FLJ36569 antibody
- MGC3245 antibody
- Msi2 antibody
see all
Images
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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 kDaLanes 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.
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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) 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.
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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 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.
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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).
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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.
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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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (34)
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