Anti-Elongation factor 1-gamma antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Elongation factor 1-gamma antibody (AB72368)

IHC image of ab72368 staining in human normal testis 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 ab72368, 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Elongation factor 1-gamma antibody (AB72368)

ICC image of ab72368 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab72368, 1 μg/mL) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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 1.43 μM.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Elongation factor 1-gamma antibody (AB72368)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma tissue labelling Elongation factor 1-gamma with ab72368 at 1/200 (1 μg/ml). Detection : DAB.

• IP

Unknown

Immunoprecipitation - Anti-Elongation factor 1-gamma antibody (AB72368)

Immunoprecipitation/ Western Blot of Elongation factor 1-gamma. Lane 1 : ab72368 at 3μg/mg whole cell lysate. Lane 2 : Control IgG. ab72368 at 1μg/ml for WB. Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded. Chemiluminescence with an exposure time of 30 seconds.

All lanes:

Immunoprecipitation - Anti-Elongation factor 1-gamma antibody (ab72368)

Predicted band size: 50 kDa

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• WB

Unknown

Western blot - Anti-Elongation factor 1-gamma antibody (AB72368)

All lanes:

Western blot - Anti-Elongation factor 1-gamma antibody (ab72368) at 0.04 µg/mL

Lane 1:

Whole cell lysate from Hela cells at 50 µg

Lane 2:

Whole cell lysate from Hela cells at 15 µg

Lane 3:

Whole cell lysate from Hela cells at 5 µg

Lane 4:

Whole cell lysate from 293T cells at 50 µg

Lane 5:

Whole cell lysate from NIH3T3 cells at 50 µg

Predicted band size: 50 kDa

Observed band size: 50 kDa

true

Exposure time: 30s

• WB

CiteAb

Western blot - Anti-Elongation factor 1-gamma antibody (AB72368)

Elongation factor 1-gamma western blot using anti-Elongation factor 1-gamma antibody ab72368. Publication image and figure legend from Ayre, D. C., Chute, I. C., et al., 2017, Sci Rep, PubMed 28819186.

ab72368 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 ab72368 please see the product overview.

Proteins identified by proteomics analysis of B cell MVs are detectable in both cell lysates and MVs of isotype and anti-CD24 stimulated B cells by Western blot analysis. Cell lysates (5 µg; equivalent to approximately 2.3 × 105 cells) and the corresponding protein from MVs (from 1.0 × 106 cells) were analyzed for expression of SHMT2, EEF1G, GRB2 and HMGB2. HSP90 was used as a loading control after stimulation with isotype (Iso) or anti-CD24 (CD24). n = 5. Two different outcomes were observed for HMGB2, with MVs from CD24 stimulated cells containing high HMGB2 (Upper panels, n = 2) or HMGB2 being low/absent in all vesicles (lower panels n = 3).

false