Anti-eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 antibody [EPR9471] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 antibody [EPR9471] - BSA and Azide free (AB240142)

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 with purified ab157455 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157455).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 antibody [EPR9471] - BSA and Azide free (AB240142)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 with purified ab157455 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157455).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 antibody [EPR9471] - BSA and Azide free (AB240142)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 with purified ab157455 at 1/50 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157455).

• IP

Unknown

Immunoprecipitation - Anti-eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 antibody [EPR9471] - BSA and Azide free (AB240142)

ab157455 (purified) at 1/30 immunoprecipitating eEF1A1/EF-Tu in HeLa whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) was used for detection at 1/1,500 dilution. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157455).

All lanes:

Immunoprecipitation - Anti-eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 antibody [EPR9471] (<a href='/en-us/products/primary-antibodies/eef1a1-ef-tu-eef1a1-ee1al3-eef1a1p5-antibody-epr9471-ab157455'>ab157455</a>)

false

• WB

Unknown

Western blot - Anti-eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 antibody [EPR9471] - BSA and Azide free (AB240142)

Exposure time :

Lane 1 : 180 seconds

Lane 2 : 5 seconds

EE1AL3/EEF1A1P5 it's a suspected pseudogene (https : //www.ncbi.nlm.nih.gov/gene/158078). Our validation shows that this clone is able to detect Recombinant EF1AL3/EEF1A1P5 but endogenous protein expression is yet unknown.

All lanes:

Western blot - Anti-eEF1A1/EF-Tu + eEF1A1 + EE1AL3/EEF1A1P5 antibody [EPR9471] (<a href='/en-us/products/primary-antibodies/eef1a1-ef-tu-eef1a1-ee1al3-eef1a1p5-antibody-epr9471-ab157455'>ab157455</a>) at 1/1000 dilution

Lane 1:

GST tagged Recombinant Human EEF1A1 protein (Full length, 76 KDa)

Lane 2:

His tagged Recombinant Human EEF1AL3 protein (Full length, 52 KDa)

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: 50 kDa

false