Anti-Elongation factor 1-gamma antibody [EPR7200]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Elongation factor 1-gamma antibody [EPR7200] (AB124994)

ab124994 at 1/100 dilution staining Elongation factor 1-gamma in HeLa cells by Immunofluorescence.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Elongation factor 1-gamma antibody [EPR7200] (AB124994)

ab124994 at 1/250 dilution staining Elongation factor 1-gamma in paraffin-embedded Human pancreatic adenocarcinoma tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Elongation factor 1-gamma antibody [EPR7200] (AB124994)

Overlay histogram showing HeLa cells stained with ab124994 (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 (ab124994 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• WB

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Western blot - Anti-Elongation factor 1-gamma antibody [EPR7200] (AB124994)

All lanes:

Western blot - Anti-Elongation factor 1-gamma antibody [EPR7200] (ab124994) at 1/10000 dilution

Lane 1:

293T cell lysate at 10 µg

Lane 2:

HL-60 cell lysate at 10 µg

Lane 3:

C6 cell lysate at 10 µg

Lane 4:

RAW 264.7 cell lysate at 10 µg

Lane 5:

PC-12 cell lysate at 10 µg

Lane 6:

NIH/3T3 cell lysate at 10 µg

Lane 7:

HepG2 cell lysate at 10 µg

Lane 8:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP-conjugated at 1/2000 dilution

Predicted band size: 50 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-Elongation factor 1-gamma antibody [EPR7200] (AB124994)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• WB

CiteAb

Western blot - Anti-Elongation factor 1-gamma antibody [EPR7200] (AB124994)

Elongation factor 1-gamma western blot using anti-Elongation factor 1-gamma antibody [EPR7200] ab124994. Publication image and figure legend from Milone, M. R., Pucci, B., et al., 2015, Oncotarget, PubMed 25481874.

ab124994 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 ab124994 please see the product overview.

Validation by 1-D Western blot of protein identified as differentially expressed in the 2-DE DIGE/MS analysisIn the middle of each validation images 1D Western blot experiment was showed. Protein lysate from DU145R80 and DU145 were immunostained with anti-LAMIN A/C (A); anti-LAMIN B2 (B); anti-AXNA1 (C); anti-FLNA (D), anti-PSMA6 (E) and anti-eEF1γ antibodies (F). CDK4 or α-tubulin immunoblot ensured equal loading of sample in each lane. The quantization of the bands was obtained using the software ImageQuantTL. Below each image the value relative to each band normalized on loading control is reported. On the left of each images the 2-DE DIGE quantification for spots of interest is reported in a 3D view that shows significant difference in protein expression. On the right the corresponding between log of standard abundance of the spot of interest (y-axis) for the two different cell lines (x-axis) on the four replicates are shown.

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