Invitrogen

Musashi-1 Monoclonal Antibody (14H1), eBioscience™

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This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

13 Published Figures

6 References

View all (39) MSI1 antibodies

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Protocols

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Musashi-1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Musashi-1 was performed using 70% confluent log phase Neural Stem cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled with Musashi-1 Monoclonal Antibody (14H1), eBioscience™ (Product # 14-9896-82) at 1 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11006), (1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e represents Neural Stem Cells differentiated to Astrocytes having no expression of Musashi-1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Musashi-1 Antibody in Western Blot (WB) — Western blot was performed using Anti-Musashi-1 Monoclonal Antibody (14H1), eBioscience™ (Product # 14-9896-80) and a 40 kDa band corresponding to Musashi-1 was observed in Neural Stem Cells and IMR-32 and not in Neural Stem Cells differentiated to Astrocytes and MCF 10A. Whole cell extracts (30 µg lysate) of Neural Stem Cells (Lane 1), Neural Stem Cells differentiated to Astrocytes (Lane 2), IMR-32 (Lane 3) and MCF 10A (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (5 ug/ml) and detected by chemiluminescence with F(ab)2-Rabbit anti-Rat IgG (H+L) Secondary Antibody, HRP (Product # PA1-29927), using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Musashi-1 Antibody — FIGURE 4. MSI1 negatively regulates NUMB and p21 expression. A, Western blotting analysis of MSI1, NUMB, and p21 in HEK-293 cells transfected with either pCMV or pCMV MSI1 plasmid at 24 h. B, mRNA levels of NUMB and p21 in pCMV or pCMV MSI1 plasmid-transfected HEK-293 cells as measured by qPCR. qPCR data were normalized to GAPDH and presented as mean S.E. of -fold changes over control, *, p < 0.05. C, Western blotting analysis of MSI1, NUMB, and p21 in HPTECs transfected with either scramble or MSI1 siRNA at 24 h. D, mRNA levels of MSI1, NUMB, and p21 in scramble and MSI1 siRNA-transfected HPTECs as measured by qPCR, qPCR data were normalized to GAPDH and presented as mean S.E. of -fold changes over scramble, *, p < 0.05. E, cell cycle analysis by propidium iodide staining and flow cytometry in HEK-293 cells transfected with either pCMV or pCMV MSI1 plasmid. F, cell cycle analysis by propidium iodide staining and flow cytometry in HPTECs transfected with either scramble or MSI1 siRNA. G, Western blotting analysis of MSI1 in HEK-293 cells transfected with either pCMV or pCMV MSI1 plasmid. H, [35S]methionine incorporation levels in pCMV or pCMV MSI1 plasmid-transfected HEK-293 cells. {KD}

Musashi-1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Extended Data Figure 1 The Musashi genes MSI1 and MSI1 are expressed in human pancreatic adenocarcinoma ( a, top row ) Representative images of a primary patient pancreatic adenocarcinoma sample stained with anti-keratin (green), DAPI (blue), and anti-MSI1 (red) antibodies. White arrows indicate MSI1 negative cells; yellow arrow indicates a MSI1 positive cell ( a, bottom row ) Representative images of a primary patient pancreatic adenocarcinoma sample stained with anti-keratin (green), DAPI (blue), and anti-MSI2 (red) antibodies. White dotted regions indicate MSI2 negative cells while yellow dotted regions indicate MSI2 positive cells. ( b, top row ) Representative images of a primary patient pancreatic adenocarcinoma sample stained with anti-keratin (green), DAPI (blue), and anti-MSI1 (red) antibodies. White arrows indicate MSI1 negative cells; yellow arrow indicates a MSI1 positive cell. ( b, bottom row ) Representative images of a primary patient pancreatic adenocarcinoma sample stained with anti-keratin (green), DAPI (blue), and anti-MSI2 (red) antibodies. Yellow dotted region indicates MSI2 positive cells. ( c, top row ) Representative images of a matched liver metastasis from a patient with pancreatic adenocarcinoma stained with anti-keratin (green), DAPI (blue), and anti-MSI1 (red) antibodies. White arrows indicate MSI1 negative cells; yellow arrows indicate MSI1 positive cells. ( c, bottom row ) Representative images of a matched liver metastasis from a patient with p

Musashi-1 Antibody in Immunohistochemistry, Immunohistochemistry (Paraffin) (IHC, IHC (P)) — Extended Data Figure 5 Selection for escaper Msi expressing cells in Msi1, Ms2 single and double knockout KP f/f C mice (a-c) Immunohistochemical staining for (a) IgG control (n=4), or (b-c, red ) Msi2 in 13 week old WT-KP f/f C (n=4) and Msi2 -/- KP f/f C (n=4) mice. (d ) Immunohistochemical staining for Msi2 (red) in 22 week old Msi2 -/- KP f/f C mouse (n=1). ( e-g ) Immunohistochemical staining for ( e ) IgG control, ( f, red) Msi1 and ( g , red) Msi2 in 15-week-old Msi1 f/f Msi2 -/- double knockout KP f/f C mouse (n=1). ( h ) Survival curves of Msi1 f/f Msi2 -/- -KP f/f C (n=6) or WT-KP f/f C tumors (n=35). Source Data for all panels are available online.

Musashi-1 Antibody in Immunohistochemistry (IHC) — Fig. 5. Early differentiation occurs around the isthmus. (A) Experimental designs of EdU short-term labeling for detection of proliferating cells, and EdU long-term labeling for cell fate tracing of proliferated cells. (B) Schematic representation of section levels used for immunohistochemistry and EdU detection (level 1 and level 3 for panels C-F). The results from six horizontal levels are shown in Fig. S5 . (C) EdU detection and immunohistochemistry for Msi1 on horizontal sections at two dorsoventral levels of the lesioned brain at 0 dpl and 4 wpl, showing that Msi1 low /EdU + cells can be found predominantly in the caudoventral mesencephalon at the early regeneration stage (arrow). In contrast, caudodorsal mesencephalon maintained undifferentiated ependymoglia cells (Msi1 high /EdU + ) at the same stage. (D-F) Caudodorsal and caudoventral sections adjacent to C (4 wpl) showed that Msi1 low /EdU + cells in the caudoventral mesencephalon (C-F, arrows) expressed a pan-neural marker HuC/D (D), Pax7 (E) and/or Lim1/2 (F). In contrast, Msi1 high /EdU + undifferentiated ependymoglia cells in the caudodorsal mesencephalon expressed Pax7 (E), and did not express HuC/D (D) and Lim1/2 (F). (G) Coronal sections of the caudal mesencephalon at 2 wpl and 6 wpl, showing that EdU-labeled cells were actively proliferating around the r0 (isthmus) between 2 wpl and 6 wpl. (H,I) Coronal sections of the caudal mesencephalon at 0 dpl and 6 wpl, showing that Pax7 + (H) and Lim1/2 + (I) ea

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

5 µg/mL

View 2 publications 2 publications

Immunohistochemistry (IHC)

View 3 publications 3 publications

Immunohistochemistry (Paraffin) (IHC (P))

Assay-Dependent

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

1 µg/mL

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Amphibian, Human, Mouse

Host/Isotype

Rat / IgG2a

Class

Monoclonal

Type

Antibody

Clone

14H1

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C

RRID

AB_10598804

Product Specific Information

Description: This 14H1 monoclonal antibody recognizes human Musashi-1 (MSI-1), a 39 kDa RNA binding protein. Musashi-1 is expressed in CNS stem and progenitor cells with decreased expression in differentiating cells. Musashi-1 functions as a translational repressor, directly regulating its target proteins, Numb and p21 (CIP1). Musashi-1 has been shown to be expressed in stem cells in a variety of other tissues such as the gut, stomach, mammary gland, and hair follicles. In addition to normal tissue expression, Musashi-1 expression has been identified in cells from several tumor types, mainly pulmonary and adenocarcinoma, as well as large and small cell carcinomas. Recently, expression of Musashi-1 in ischemic lesions following stroke provides evidence for neurogenesis as well as the role of Musashi-1 in regulating apoptosis.

Musashi-1 is highly conserved and the 14H1 antibody also recognizes mouse and rat Musashi-1.

Applications Reported: This 14H1 antibody has been reported for use in immunoblotting (WB), immunohistology staining of paraffin embedded tissue sections, and immunocytochemistry.

Applications Tested: This 14H1 antibody has been tested by immunofluorescent staining of paraformaldehyde fixed and permeabilized cells. This can be used at less than or equal to 10 µg/mL. It is recommended that the antibody be titrated for optimal performance in the assay of interest.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Musashi-1 belongs to the Musashi family of RNA-binding proteins which play a role in cell fate determination, and maintenance of stem-cell state. Musashi family proteins are also involved in post transcriptional gene regulation. The gene encodes a protein with two conserved tandem RNA recognition motifs with highly conserved RNP (ribonucleoprotein) motifs. In mammals, Musashi-1 is expressed in fetal kidney, brain, liver and lung, and in adult brain and pancreas. In humans, the gene is located on chromosome 12.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.