Anti-eIF3e antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-eIF3e antibody (AB36766)

ab36766 staining eIF3e in MCF7 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 ab36766 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 confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IP

Unknown

Immunoprecipitation - Anti-eIF3e antibody (AB36766)

eIF3e was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to eIF3e 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 ab36766.
Secondary : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band : 48kDa : eIF3e.

All lanes:

Immunoprecipitation - Anti-eIF3e antibody (ab36766)

Predicted band size: 52 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF3e antibody (AB36766)

IHC image of ab36766 staining in pancreas 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 ab36766, 5µg/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.

• WB

AbReview33205****

Western blot - Anti-eIF3e antibody (AB36766)

All lanes:

Western blot - Anti-eIF3e antibody (ab36766) at 1/1000 dilution

Lane 1:

Chicken tectum lysate at 5 µg

Lane 2:

Zebrafish lysate at 5 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit polyclonal IgG at 1/10000 dilution

Predicted band size: 52 kDa

Observed band size: 38 kDa,48 kDa

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This image is courtesy of an anonymous Abreview

• WB

Project

Western blot - Anti-eIF3e antibody (AB36766)

All lanes:

Western blot - Anti-eIF3e antibody (ab36766) 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:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg with Human eIF3e peptide (<a href='/en-us/products/unavailable/human-eif3e-peptide-ab36902'>ab36902</a>)

Lane 6:

Western blot - Jurkat whole cell lysate (<a href='/en-us/products/cell-lysates/jurkat-whole-cell-lysate-ab7899'>ab7899</a>) at 10 µg with Human eIF3e peptide (<a href='/en-us/products/unavailable/human-eif3e-peptide-ab36902'>ab36902</a>)

Lane 7:

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 with Human eIF3e peptide (<a href='/en-us/products/unavailable/human-eif3e-peptide-ab36902'>ab36902</a>)

Lane 8:

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 with Human eIF3e peptide (<a href='/en-us/products/unavailable/human-eif3e-peptide-ab36902'>ab36902</a>)

Secondary

All lanes:

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

Predicted band size: 52 kDa

Observed band size: 40 kDa,48 kDa

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