Anti-Brn-2 antibody [RP23040248]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Brn-2 antibody [RP23040248] (AB308102)

For immunofluorescence analysis, Neural stem cells were fixed and permeabilized for detection of endogenous Brn-2 using ab308102 at 1/100 dilution and labeled with a Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution. Panel a) shows representative cells that were stained for detection and localization of Brn-2 protein (green) Panel b) is stained for nuclei (blue) using DAPI Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin at 1/300 dilution Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of Brn-2 Panel e) represents control cells with no primary antibody to assess background The images were captured at 60X magnification

• WB

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Western blot - Anti-Brn-2 antibody [RP23040248] (AB308102)

Knockdown of Brn-2 was achieved by transfecting A-375 cells with Brn-2 specific siRNA. Western blot analysis was performed using modified whole cell extracts (1% SDS) from the A-375 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with ab308102 at 1/200 dilution and a Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4,000 dilution. Loss of signal upon siRNA mediated knock down confirms that antibody is specific to POU3F2.

All lanes:

Western blot - Anti-Brn-2 antibody [RP23040248] (ab308102) at 1/200 dilution

Lane 1:

untransfected A375 cell lysate

Lane 2:

non-specific siRNA transfected A375 cell lysate

Lane 3:

Brn-2 specific siRNA transfected A375 cell lysate

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution

Observed band size: 54 kDa

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• WB

Supplier Data

Western blot - Anti-Brn-2 antibody [RP23040248] (AB308102)

Western blot analysis was performed on modified whole cell extracts (1%SDS) (30 µg lysate) of U-87 MG (Lane 1) and tissue extracts of Mouse pup brain (Lane 2) and Mouse brain (Lane 3). The blots were probed with ab308102 at 1/200 dilution and detected by chemiluminescence using a Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution. A ~54 kDa band corresponding to POU3F2 was observed across the cell lines and tissues tested.

All lanes:

Western blot - Anti-Brn-2 antibody [RP23040248] (ab308102) at 1/200 dilution

Lane 1:

U-87 MG cell lysate at 30 µg

Lane 2:

Mouse pup brain tissue lysate at 30 µg

Lane 3:

Mouse brain tissue lysate at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution

Observed band size: 54 kDa

false