711843

antibody from Invitrogen Antibodies

Targeting: EEF2 EEF-2, EF2

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

711843

Provider

Invitrogen Antibodies

Product name

Phospho-EEF2 (Thr56) Recombinant Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

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 Jurkat (Lane 1), T98G (Lane 2), Hep G2 (Lane 3), 3T3-L1 (Lane 4), 3T3-L1 treated with Forskolin (100uM for 60 mins) (Lane 5), NIH/3T3 (Lane 6), NIH/3T3 treated with Forskolin (100uM for 60 mins) (Lane 7). The blots were probed with Anti-EEF2 pT56 Recombinant Rabbit Polyclonal Antibody (Product # 711843, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 95 kDa band corresponding to EEF2 pT56 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on Whole cell extracts (30 µg lysate) of NIH/3T3 (Lane 1) and NIH/3T3 treated with Forskolin (100uM for 60 mins) (Lane 2). The blots were probed with Anti-EEF2 pT56 Recombinant Rabbit Polyclonal Antibody (Product # 711843, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 95 kDa band corresponding to EEF2 pT56 was observed. Antibody specificity was demonstrated by competition with the EEF2 pT56 peptide, which results in loss of signal. No competition was observed with the non-phospho peptide. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), 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).