EEF2K Recombinant Rabbit Monoclonal Antibody [PSH11-67] - BSA and Azide free

This data was developed using HA723363, the same antibody clone in a different buffer formulation.
Western blot analysis of EEF2K on different lysates with Rabbit anti-EEF2K antibody (HA723363) at 1/10,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: K-562 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 6: Mouse ovary tissue lysate (40 µg/Lane)
Lane 7: Rat ovary tissue lysate (40 µg/Lane)

Predicted band size: 82 kDa
Observed band size: 82-90 kDa

Exposure time: 25 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723363) at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

This data was developed using HA723363, the same antibody clone in a different buffer formulation.
Western blot analysis of EEF2K on different lysates with Rabbit anti-EEF2K antibody (HA723363) at 1/5,000 dilution.

Lane 1: HeLa-parental cell lysate
Lane 2: HeLa-EEF2K KD cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 82 kDa
Observed band size: 82-90 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723363) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

This data was developed using HA723363, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa cells labeling EEF2K with Rabbit anti-EEF2K antibody (HA723363) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-EEF2K antibody (HA723363) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

This data was developed using HA723363, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of HeLa cells labeling EEF2K.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723363, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).