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Anti-EEF2 Antibody EP880Y
Also for EEF2 (NM_001961)
|A synthetic peptide corresponding to residues on the C-terminus of human eEF2 was used as an immunogen.|
|Mouse, Rat, Human
||Lot dependent; please refer to CoA along with shipment
|WB, IHC, IF, IP, FC
||WB: 1:10000 - 1:20000; IP: Use a concentration of 5 ug/ml; FC: 1:20; IHC-P: 1:100 - 1:250; ICC: 1:100 - 1:250; ICC/IF: Use a concentration of 1 ug/ml
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant
|Homo sapiens eukaryotic translation elongation factor 2 (EEF2)|
|EEF-2; EF-2; EF2; SCA26|
Entrez Gene 1938 Human
Entrez Gene 13629 Mouse
Entrez Gene 29565 Rat
|This gene encodes a member of the GTP-binding translation elongation factor family. This protein is an essential factor for protein synthesis. It promotes the GTP-dependent translocation of the nascent protein chain from the A-site to the P-site of the ribosome. This protein is completely inactivated by EF-2 kinase phosporylation. |
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Western blot - EEF2 antibody [EP880Y] - Carboxyterminal end; All lanes : Anti-EEF2 antibody [EP880Y] - C-terminal at 1/10000 dilution.Lane 1 : 293 cell lysate.Lane 2 : HeLa cell lysate.Lane 3 : A431 cell lysate.Lane 4 : NIH3T3 cell lysate.Lysates/proteins at 10 ug per lane.Secondary.HRP labelled goat anti-rabbit at 1/2000 dilution.Predicted band size : 95 kDa.Observed band size : 95 kDa.Additional bands at : 30 kDa,70 kDa. We are unsure as to the identity of these extra bands.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - EEF2 antibody [EP880Y] - C-terminal; TA303826 at 1/100 dilution staining EEF2 in human kidney carcinoma tissue.
Immunocytochemistry/ Immunofluorescence-EEF2 antibody [EP880Y] - C-terminal(TA303826); ICC/IF image of TA303826 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was Alexa Fluor 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43uM.
Immunoprecipitation - Anti-EEF2 antibody [EP880Y] - C-terminal; EEF2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5ug of Rabbit monoclonal to EEF2 and 50ul of protein G magnetic beads (+). No antibody was added to the control (-). .The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40ul SDS loading buffer and incubated for 10min at 70oC; 10ul of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with TA303826.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) .Band: 95kDa; EEF2
Flow Cytometry - Anti-EEF2 antibody [EP880Y] - C-terminal; Overlay histogram showing HeLa cells stained with TA303826 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1ug/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed.