Anti-RPS6 antibody [RP23040083]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RPS6 antibody [RP23040083] (AB313428)

Immunofluorescent analysis of 70% confluent log phase U-2 OS cells fixed with 4% paraformaldehyde for 15 minutes, and blocked with 5% BSA for 1 hour at room temperature labeling RPS6 with ab313428 at 1 ?g/mL, followed by Alexa Fluor 488 Goat anti-Rabbit IgG Secondary Antibody at a dilution of 1/400 for 30 minutes at room temperature (Panel a/ green). Nuclei (Panel b/ blue) were stained with DAPI. F-actin (Panel c/ red) was stained with Alexa Fluor 594 Phalloidin. Panel d is a merged image showing cytoplasmic localization and panel e is a no primary antibody control. The images were captured at 20X magnification.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RPS6 antibody [RP23040083] (AB313428)

Immunofluorescent analysis of HeLa cells fixed using 4% formaldehyde (reconstituted in 1X PBS) for 10 min at room temperature and permeabilized using 0.1 % Triton X-100 in PBS for 15 min at room temperature labeling RPS6 with ab313428 followed by detection using an Alexa Fluor 488-conjugated Goat anti-Rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing cytoplasmic localization of RPS6.

• Flow Cyt

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Flow Cytometry - Anti-RPS6 antibody [RP23040083] (AB313428)

Flow cytometry analysis of FIX & PERM reagent fixed Hela cells labeling RPS6 with ab313428. Detection was performed using an Alexa Fluor 488 Goat anti-Rabbit IgG (right peak) compared to an isotype control (left peak).

• IP

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Immunoprecipitation - Anti-RPS6 antibody [RP23040083] (AB313428)

RPS6 was immunoprecipitated from HeLa cells whole cell lysate with ab313428 at 2 µg/mL. Western blot was performed from the immunoprecipitate using ab313428 at 1 µg/mL. Rabbit IgG was used as a IP control.

All lanes:

Immunoprecipitation - Anti-RPS6 antibody [RP23040083] (ab313428) at 1 µg/mL

Lane 1:

Immunoprecipitated HeLa cell lysate

Lane 2:

Rabbit IgG control

Predicted band size: 29 kDa

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• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RPS6 antibody [RP23040083] (AB313428)

Immunofluorescent analysis of 3T3 cells permeabilized with 0.1% Triton X-100 in TBS for 15 minutes, and blocked with 3% Blocker BSA in PBS for 15 minutes at room temperature labeling S6 ribosomal protein (green) with ab313428 at 10 ?g/mL in blocking buffer for at least 1 hour at room temperature, and then incubated with a Goat anti-Rabbit IgG secondary antibody, Alexa Fluor 488 conjugate at a dilution of 1/1000 for 30 minutes at room temperature (green). Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

• WB

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Western blot - Anti-RPS6 antibody [RP23040083] (AB313428)

All lanes:

Western blot - Anti-RPS6 antibody [RP23040083] (ab313428) at 0.5 µg/mL

Lane 1:

MCF7 cell lysate at 30 µg

Lane 2:

HEK-293 cell lysate at 30 µg

Lane 3:

Hep G2 cell lysate at 30 µg

Lane 4:

Jurkat cell lysate at 30 µg

Lane 5:

SK-N-SH cell lysate at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG-HRP Secondary Antibody at 1/5000 dilution

Predicted band size: 28.681 kDa

Observed band size: 31 kDa

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• WB

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Western blot - Anti-RPS6 antibody [RP23040083] (AB313428)

All lanes:

Western blot - Anti-RPS6 antibody [RP23040083] (ab313428) at 1/1000 dilution

Lane 1:

Untransfected HeLa cell lysate

Lane 2:

non-specific scrambled siRNA transfected HeLa cell lysate

Lane 3:

S6 HeLa knockdown cell lysate

Secondary

All lanes:

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

Predicted band size: 28.681 kDa

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• WB

Supplier Data

Western blot - Anti-RPS6 antibody [RP23040083] (AB313428)

All lanes:

Western blot - Anti-RPS6 antibody [RP23040083] (ab313428) at 1 µg/mL

Lane 1:

TNF alpha treated HeLa cell lysate

Lane 2:

Untreated HeLa cell lysate

Lane 3:

A431 cell lysate

Lane 4:

HepG2 cell lysate

Lane 5:

Jurkat cell lysate

Lane 6:

HEK293 cell lysate

Lane 7:

MCF7 cell lysate

Lane 8:

PC12 cell lysate

Predicted band size: 28.681 kDa

Observed band size: 31 kDa

true