- PA5-27449 - Invitrogen Antibodies MSI1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of MSI1 using 30 µg of H1299 lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a MSI1 polyclonal antibody (Product # PA5-27449) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of MSI1 was performed by separating 50 µg of various tissue extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a MSI1 Polyclonal Antibody (Product # PA5-27449) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of MSI1 was performed by separating 50 µg of various tissue extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a MSI1 Polyclonal Antibody (Product # PA5-27449) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of MSI1 was performed by separating 30 µg of whole cell extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a MSI1 Polyclonal Antibody (Product # PA5-27449) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of MSI1 was achieved by transfecting HEK-293 with MSI1 specific siRNAs (Silencer® select Product # s8980; s8979). Western blot analysis (Fig. a) was performed using whole cell extracts from the MSI1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with MSI1 Polyclonal Antibody (Product # PA5-27449, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to MSI1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), PC-12 (Lane 2) and HEK-293 (Lane 3). The blot was probed with Anti-MSI1 Polyclonal Antibody (Product # PA5-27449, 1:2000 dilution) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 40kDa band corresponding to MSI1 was observed across all the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of MSI1 in paraformaldehyde-fixed A549 cells using a MSI1 polyclonal antibody (Product # PA5-27449) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of MSI1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: MSI1 Polyclonal Antibody (Product # PA5-27449) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of MSl1 was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with anti-MSI1 Polyclonal Antibody (Product # PA5-27449) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). 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 control cells with no primary antibody to assess background. The images were captured at 60X magnification

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Frozen) analysis of MSI1 was performed in frozen sectioned adult mouse retina tissue using MSI1 Polyclonal Antibody (Product # PA5-27449) at a dilution of 1:250 (Green). Red: Protein kinase C alpha staining. Blue: Fluoroshield with DAPI. Antigen Retrieval: EDTA based buffer, pH 8.0 buffer, 15min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: MSI1 antibody [N3C3] detects MSI1 protein on adult mouse brain by immunohistochemical analysis. Sample: Frozen section of adult mouse brain. MSI1 antibody [N3C3] (Product # PA5-27449) diluted at 1:500.

PA5-27449