Anti-IRS1 (phospho S312) antibody [RP23040108]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IRS1 (phospho S312) antibody [RP23040108] (AB313437)

Immunofluorescent analysis of MCF7 cells treated with IGF (100 ng/mL/20 min) 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 IRS with ab313437 at 1 ?g/mL followed by Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 488 conjugate at 1/2500 dilution. Panel a) shows representative cells that were stained for detection and localization of IRS (pS312) protein (green), Panel b) is stained for Nuclei (blue) using DAPI at 1/50 dilution. Panel c) represents cytoskeletal F-actin staining using Alexa Fluor 594 Phalloidin at 1/200 dilution. Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of IRS (pS312). Panel e) represents cells without treatment and panel f) represents control cells with no primary antibody to assess background.

• WB

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Western blot - Anti-IRS1 (phospho S312) antibody [RP23040108] (AB313437)

All lanes:

Western blot - Anti-IRS1 (phospho S312) antibody [RP23040108] (ab313437) at 1 µg/mL

Lane 1:

HepG2 cell lysate at 30 µg

Lane 2:

HepG2 treated with IGF1 (100 ng/mL/20 min) cell lysate at 30 µg

Lane 3:

MCF7 cell lysate at 30 µg

Lane 4:

MCF7 treated with Insulin (100 nM/10 min) cell lysate at 30 µg

Lane 5:

C2C12 cell lysate at 30 µg

Lane 6:

C2C12 treated with Insulin (100 nM/ 10 minutes) cell lysate at 30 µg

Lane 7:

phosphopeptide incubated (10 µg/ml) HepG2 cell lysate at 30 µg

Lane 8:

phosphopeptide incubated (10 µg/ml) HepG2 treated with IGF1 (100 ng/mL/20 min) cell lysate at 30 µg

Lane 9:

phosphopeptide incubated (10 µg/ml) MCF7 cell lysate at 30 µg

Lane 10:

phosphopeptide incubated (10 µg/ml) MCF7 treated with Insulin (100 nM/10 min) cell lysate at 30 µg

Lane 11:

phosphopeptide incubated (10 µg/ml) C2C12 cell lysate at 30 µg

Lane 12:

phosphopeptide incubated (10 µg/ml) C2C12 treated with Insulin (100 nM/10 min) cell lysate at 30 µg

Secondary

All lanes:

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

Predicted band size: 131.591 kDa

Observed band size: 180 kDa

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