710688

antibody from Invitrogen Antibodies

Targeting: FOS AP-1, c-fos

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

710688

Provider

Invitrogen Antibodies

Product name

Phospho-c-Fos (Thr232) Recombinant Polyclonal Antibody (3HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

3HCLC

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

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 Anti-c-Fos (pT232) Recombinant Rabbit Polyclonal Antibody (Product # 710688, 2-3 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A clear 40 kDa band corresponding to c-Fos (pT232) was observed across cell lines tested. B) To confirm the specificity of Anti-c-Fos (pT232) Recombinant Rabbit Polyclonal Antibody, competition was performed with the phosphopeptide (10 µg/mL) as shown in the corresponding blot on right. The peptide competes with the antibody and prevents it from binding to the membrane. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).