Anti-HJURP antibody [RP23040269]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HJURP antibody [RP23040269] (AB308103)

For immunofluorescence analysis, A549 cells were fixed and permeabilized for detection of endogenous HJURP using ab308103 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 HJURP 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 HJURP 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-HJURP antibody [RP23040269] (AB308103)

Western blot analysis was performed on modified whole cell extracts (30 µg lysate) of HeLa (Lane 1), T98G (Lane 2), U-87 MG (Lane 3) and U-2 OS (Lane 4). The blot was probed with ab308103 at 1 : 500 dilution and detected by chemiluminescence using a Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1 : 4000 dilution. A ~83 kDa band corresponding to HJURP was observed across the cell lines tested.

All lanes:

Western blot - Anti-HJURP antibody [RP23040269] (ab308103) at 1/500 dilution

Lane 1:

HeLa whole cell lysate at 30 µg

Lane 2:

T98G whole cell lysate at 30 µg

Lane 3:

U-87 MG whole cell lysate at 30 µg

Lane 4:

U-2 OS whole cell lysate at 30 µg

Secondary

All lanes:

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

Observed band size: 83 kDa

true

• WB

Supplier Data

Western blot - Anti-HJURP antibody [RP23040269] (AB308103)

Knockdown of HJURP was achieved by transfecting A549 cells with HJURP specific siRNA. Western blot analysis was performed using modified whole cell extracts (1% SDS) from HJURP knockdown cells (Lane 2) and non-specific scrambled siRNA transfected cells (Lane 1). The blot was probed with ab308103 at 1 : 500 dilution and a Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1 : 4000 dilution. Loss of signal upon siRNA mediated knockdown confirms that antibody is specific to HJURP.

All lanes:

Western blot - Anti-HJURP antibody [RP23040269] (ab308103) at 1/500 dilution

Lane 1:

non-specific scrambled siRNA transfected A549 cells

Lane 2:

HJURP knockdown A549 cells

Secondary

All lanes:

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

Observed band size: 83 kDa

false