Anti-EEF2/Elongation factor 2 (phospho T56) antibody [RP23040245]

• WB

Supplier Data

Western blot - Anti-EEF2/Elongation factor 2 (phospho T56) antibody [RP23040245] (AB308101)

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 ab308101 and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1 : 4000 dilution. A 95 kDa band corresponding to EEF2 (phospho T56) was observed across the 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-EEF2/Elongation factor 2 (phospho T56) antibody [RP23040245] (ab308101) at 2.5 µg/mL

Lane 1:

Jurkat cell lysate at 30 µg

Lane 2:

T98G cell lysate at 30 µg

Lane 3:

HepG2 cell lysate at 30 µg

Lane 4:

3T3-L1 cell lysate at 30 µg

Lane 5:

Forskolin treated 3T3-L1 cell lysate at 30 µg

Lane 6:

NIH/3T3 cell lysate at 30 µg

Lane 7:

Forskolin treated NIH/3T3 cell lysate at 30 µg

Secondary

All lanes:

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

Observed band size: 95 kDa

true

• WB

Supplier Data

Western blot - Anti-EEF2/Elongation factor 2 (phospho T56) antibody [RP23040245] (AB308101)

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 ab308101 and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1 : 4000 dilution. A 95 kDa band corresponding to EEF2 (phospho T56) was observed. Antibody specificity was demonstrated by competition with the EEF2 (phospho T56) peptide, which results in loss of signal. No competition was observed with the non-phospho peptide. 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-EEF2/Elongation factor 2 (phospho T56) antibody [RP23040245] (ab308101) at 2.5 µg/mL

Lane 1:

untreated NIH/3T3 cell lysate at 30 µg

Lane 2:

Forskolin treated NIH/3T3 cell lysate at 30 µg

Lane 3:

EEF2 (phospho T56) phosphopeptide incubated NIH/3T3 cell lysate at 30 µg

Lane 4:

EEF2 (phospho T56) phosphopeptide incubated Forskolin treated NIH/3T3 cell lysate at 30 µg

Lane 5:

non-phosphopeptide incubated NIH/3T3 cell lysate at 30 µg

Lane 6:

Non-phosphopeptide treated Forskolin treated NIH/3T3 cell lysate at 30 µg

Secondary

All lanes:

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

Observed band size: 95 kDa

true