Anti-RIP Antibody [RP23040227]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RIP Antibody [RP23040227] (AB313489)

Immunofluorescent analysis of HT29 cells transfected with RIPK1 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 RIPK1 with ab313489 at 5 ?g/mL followed by Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 488 conjugate at 1/2000 dilution. Panel a-d) untransfected HT29 cells. Panel b, e) HT29 cells transfected with non-specific scrambled siRNA. Panel c,f) HT29 cells transfected with RIPK1 specific siRNA. Nuclei (blue) were stained using DAPI, and Rhodamine Phalloidin at 1/300 dilution. was used for cytoskeletal F-actin (red) staining. Loss of signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to RIPK1 (green). The images were captured at 60X magnification

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RIP Antibody [RP23040227] (AB313489)

Immunofluorescent analysis of COLO-205 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 RIPK1 with ab313489 at 5 ?g/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 RIPK1 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 cytoplasmic localization of RIPK1. Panel represents control cells with no primary antibody to assess background. Panel f) represents SW480 cells which is negative for RIPK1 showing no signal.

• WB

Supplier Data

Western blot - Anti-RIP Antibody [RP23040227] (AB313489)

All lanes:

Western blot - Anti-RIP Antibody [RP23040227] (ab313489) at 2.5 µg/mL

Lane 1:

Jurkat cell lysate at 30 µg

Lane 2:

COLO205 cell lysate at 30 µg

Lane 3:

SW480 cell lysate at 30 µg

Lane 4:

HT29 cell lysate at 30 µg

Lane 5:

HCT116 cell lysate at 30 µg

Lane 6:

MDA-MB-231 cell lysate at 30 µg

Lane 7:

MCF7 cell lysate at 30 µg

Lane 8:

HepG2 cell lysate at 30 µg

Lane 9:

A549 cell lysate at 30 µg

Lane 10:

HeLa cell lysate at 30 µg

Secondary

All lanes:

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

Observed band size: 76 kDa

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