Anti-RME-8 antibody [RP23040241]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RME-8 antibody [RP23040241] (AB313497)

Immunofluorescent analysis of U-87 MG cells fixed with 4% formaldehyde reconstituted in 1X PBS for 10 min at room temperature and permabilized using 0.1 % Triton X-100 in PBS for 15 min at room temperature labeling endogenous DNAJC13 with ab313497 at 2 µg/mL dilution followed by 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 DNAJC13 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 endosome localization of RME-8. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RME-8 Antibody [RP23040241] (AB313497)

Immunofluorescent analysis of U-87 MG cells transfected with specific siRNA fixed with 4% formaldehyde reconstituted in 1X PBS for 10 min at room temperature and permabilized using 0.1 % Triton X-100 in PBS for 15 min at room temperature labeling DNAJC13 with ab313497 at 1/100 dilution followed by Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 488 conjugate at 1/2000 dilution. Panel a-d) untransfected U-87MG cells. Panel i-l) U-87MG cells transfected with DNAJC13 specific siRNA. Panel e-h) U-87MG cells transfected with non-specific scrambled siRNA. Nuclei (blue) were stained using DAPI, and Rhodamine Phalloidin at 1/300 dilution was used for cytoskeletal F-actin (red) staining. Significant reduction of signal was observed upon siRNA mediated knockdown (panel i-l) confirming specificity of the antibody to DNAJC13 (green). The images were captured at 60X magnification.

• WB

Supplier Data

Western blot - Anti-RME-8 Antibody [RP23040241] (AB313497)

All lanes:

Western blot - Anti-RME-8 Antibody [RP23040241] (ab313497) at 1 µg/mL

Lane 1:

COS7 cell lysate at 30 µg

Lane 2:

U2OS cell lysate at 30 µg

Lane 3:

MDA-MB-231 cell lysate at 30 µg

Lane 4:

MCF7 cell lysate at 30 µg

Lane 5:

T47D cell lysate at 30 µg

Lane 6:

HepG2 cell lysate at 30 µg

Lane 7:

HEK293 cell lysate at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/2500 dilution

Observed band size: 254 kDa

true

• WB

Supplier Data

Western blot - Anti-RME-8 Antibody [RP23040241] (AB313497)

All lanes:

Western blot - Anti-RME-8 Antibody [RP23040241] (ab313497) at 1 µg/mL

Lane 1:

untransfected MCF7 cell lysate

Lane 2:

non-specific scrambled siRNA transfected MCF7 cell lysate

Lane 3:

RME8 specific siRNA transfected MCF7 cell lysate

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/5000 dilution

Observed band size: 228 kDa

false