Anti-c-Fos (phospho T232) antibody [RP23040102]

• WB

Supplier Data

Western blot - Anti-c-Fos (phospho T232) antibody [RP23040102] (AB308128)

Blot B. To confirm the specificity of ab308128, competition was performed with the phosphopeptide (10 µg/mL) as shown. The peptide competes with the antibody and prevents it from binding to the membrane. Known quantity of protein samples were electrophoresed using a 4-12% Bis-Tris gel, electrophoresis system and pre-stained protein standard. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed (ECL).

All lanes:

Western blot - Anti-c-Fos (phospho T232) antibody [RP23040102] (ab308128) at 2 µg/mL

Lane 1:

HeLa cell lysate with phosphopeptide at 30 µg

Lane 2:

HeLa cell lysate treated with EGF and phosphopeptide at 30 µg

Lane 3:

A431 cell lysate with phosphopeptide at 30 µg

Lane 4:

A431 cell lysate treated with EGF and phosphopeptide at 30 µg

Secondary

All lanes:

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

Predicted band size: 41 kDa

true

• WB

Supplier Data

Western blot - Anti-c-Fos (phospho T232) antibody [RP23040102] (AB308128)

Blot A. Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (lane 1), HeLa treated with EGF (200 µg/mL/10 min), A-431 (lane 3) and A-431 treated with EGF (200 µg/mL/10 min) (lane 4). The blots were probed with ab308128, 2-3 µg/mL and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (0.4 µg/mL, 1 : 2500 dilution). A clear 40 kDa band corresponding to c-Fos (phospho T232) was observed across cell lines tested. Known quantity of protein samples were electrophoresed using a 4-12% Bis-Tris gel, electrophoresis system and pre-stained protein standard. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed (ECL).

All lanes:

Western blot - Anti-c-Fos (phospho T232) antibody [RP23040102] (ab308128) at 2 µg/mL

Lane 1:

HeLa cell lysate at 30 µg

Lane 2:

HeLa cell lysate treated with EGF (200 µg/mL/10 min) at 30 µg

Lane 3:

A431 cell lysate at 30 µg

Lane 4:

A-431 cell lysate treated with EGF (200 µg/mL/10 min) at 30 µg

Secondary

All lanes:

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

Predicted band size: 41 kDa

Observed band size: 40 kDa

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