Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free

• WB

Lab

Western blot - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal[EPR24714-88] to EEF2K ab270948 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 82 kDa in Wild-type A549 cell lysates with no signal observed at this size in EEF2K knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

Lanes 1 - 4:

Western blot - Anti-EEF2K antibody [EPR24714-88] (<a href='/en-us/products/primary-antibodies/eef2k-antibody-epr24714-88-ab270948'>ab270948</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (ab283706) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

EEF2K knockout A549 at 20 µg

Lane 3:

Raji at 20 µg

Lane 4:

MOLT-4 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 82 kDa

Observed band size: 82 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labelling EEF2K with ab270948 at 1/500 (1.178 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human pancreatic acinar cells. The section was incubated with ab270948 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling EEF2K with ab270948 at 1/500 (1.178 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human breast carcinoma. The section was incubated with ab270948 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling EEF2K with ab270948 at 1/500 (1.178 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : no staining on human liver. The section was incubated with ab270948 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling EEF2K with ab270948 at 1/50 dilution (1ug)/(Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labelling EEF2K with ab270948 at 1/500 (1.178 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human endometrial carcinoma. The section was incubated with ab270948 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) cells labelling EEF2K with ab270948 at 1/50 dilution (1ug)/ (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

EEF2K was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab270948 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270948 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab270948 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270948 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 32 seconds

All lanes:

Immunoprecipitation - Anti-EEF2K antibody [EPR24714-88] (<a href='/en-us/products/primary-antibodies/eef2k-antibody-epr24714-88-ab270948'>ab270948</a>)

Predicted band size: 82 kDa

false

• WB

Lab

Western blot - Anti-EEF2K antibody [EPR24714-88] - BSA and Azide free (AB283706)

This data was developed using ab270948, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Lysates were made freshly and used in WB test immediately to minimize protein degradation.

Exposure time : 10 seconds

All lanes:

Western blot - Anti-EEF2K antibody [EPR24714-88] (<a href='/en-us/products/primary-antibodies/eef2k-antibody-epr24714-88-ab270948'>ab270948</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 82 kDa

Observed band size: 95 kDa

false