Anti-EEF2/Elongation factor 2 antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-EEF2/Elongation factor 2 antibody (AB33523)

ab33523 staining EEF2 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab33523 at 1μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EEF2/Elongation factor 2 antibody (AB33523)

IHC image of EEF2/Elongation factor 2 staining in human lung formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab33523, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IP

Unknown

Immunoprecipitation - Anti-EEF2/Elongation factor 2 antibody (AB33523)

EEF2/Elongation factor 2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5μg of Rabbit polyclonal to EEF2/Elongation factor 2 and 50μl 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 40μl SDS loading buffer and incubated for 10min at 70^oC; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab33523.
Secondary : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band : 95kDa; EEF2/Elongation factor 2, non-specific band : 135kDa,due to background seen in No ab control lane (2).

All lanes:

Immunoprecipitation - Anti-EEF2/Elongation factor 2 antibody (ab33523)

Predicted band size: 95 kDa

false

• WB

Project

Western blot - Anti-EEF2/Elongation factor 2 antibody (AB33523)

All lanes:

Western blot - Anti-EEF2/Elongation factor 2 antibody (ab33523) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Western blot - Jurkat whole cell lysate (<a href='/en-us/products/cell-lysates/jurkat-whole-cell-lysate-ab7899'>ab7899</a>) at 10 µg

Lane 3:

Western blot - A-431 whole cell lysate (<a href='/en-us/products/cell-lysates/a-431-whole-cell-lysate-ab7909'>ab7909</a>) at 10 µg

Lane 4:

Western blot - HEK-293 whole cell lysate (<a href='/en-us/products/cell-lysates/hek-293-whole-cell-lysate-ab7902'>ab7902</a>) at 10 µg

Lane 5:

Hep G2 whole cell lysate (<a href='/en-us/products/unavailable/hep-g2-whole-cell-lysate-ab7900'>ab7900</a>) at 10 µg

Lane 6:

MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 7:

SHSY-5Y (Human Neuroblastoma) Whole Cell Lysate at 10 µg

Lane 8:

U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

Predicted band size: 95 kDa

Observed band size: 95 kDa

false

• WB

Unknown

Western blot - Anti-EEF2/Elongation factor 2 antibody (AB33523)

All lanes:

Western blot - Anti-EEF2/Elongation factor 2 antibody (ab33523) at 1 µg/mL

Lane 1:

Western blot - NIH/3T3 whole cell lysate (<a href='/en-us/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 10 µg

Lane 2:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 3:

Western blot - Mouse pancreas tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-pancreas-tissue-lysate-total-protein-ab29363'>ab29363</a>) at 10 µg

Lane 4:

Ovary (Mouse) Tissue Lysate - normal tissue at 10 µg

Lane 5:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Lane 6:

Liver (Rat) Tissue Lysate - normal tissue at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 95 kDa

Observed band size: 95 kDa

false

• WB

Unknown

Western blot - Anti-EEF2/Elongation factor 2 antibody (AB33523)

All lanes:

Western blot - Anti-EEF2/Elongation factor 2 antibody (ab33523) at 1 µg/mL

Lane 1:

Marker

Lane 2:

Zebrafish brain homogenate (20ug)

Lane 3:

Zebrafish heart homogenate (20ug)

Lane 4:

Zebrafish liver homogenate (20ug)

Lane 5:

Zebrafish skeletal muscle homogenate (20ug)

Lane 6:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (20ug)

Secondary

All lanes:

Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

Predicted band size: 95 kDa

Observed band size: 95 kDa

true

Exposure time: 5min

• WB

CiteAb

Western blot - Anti-EEF2/Elongation factor 2 antibody (AB33523)

EEF2/Elongation factor 2 western blot using anti-EEF2/Elongation factor 2 antibody ab33523. Publication image and figure legend from Abere, B., Wikan, N., et al., 2012, PLoS One, PubMed 22514668.

ab33523 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab33523 please see the product overview.

Analysis of CHIKV infected CHME-5 cells.CHME-5 cells either mock infected or infected with CHIKV at MOI 0.1 were collected at day 2 p.i. (A, B, D) or on days 1 to 3 p.i. (C) and subsequently (A) stained with an anti-alphavirus antibody and the percentage of infected cells analyzed by flow cytometry or (B) stained with Annexin V-FITC and PI and the percentage of apoptotic cells analyzed by flow cytometery or (C, D) used for total protein extraction and (C) analyzed by western blot with an anti-alphavirus monoclonal antibody and an anti-actin polyclonal antibody or (D) the differential proteome determined by 2D-PAGE. Representative gels from 6 biological replicates are shown. (A and B) Bar graphs represent the means ± SD of 6 replications.

false