Anti-eIF3e antibody (ab36766)
Key features and details
- Rabbit polyclonal to eIF3e
- Suitable for: IHC-P, ICC/IF, WB, IP
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-eIF3e antibody
See all eIF3e primary antibodies -
Description
Rabbit polyclonal to eIF3e -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIP HumanWB Human
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human eIF3e.
Read Abcam's proprietary immunogen policy (Peptide available as ab36902.) -
Positive control
- Recombinant Human eIF3e protein (ab114769) can be used as a positive control in WB. This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line) A431 (Human epithelial carcinoma cell line) HEK 293 (Human embryonic kidney cell line) HepG2 (Human hepatocellular liver carcinoma cell line) MCF-7 (Human breast adenocarcinoma cell line) SHSY-5Y (Human neuroblastoma cell line)
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Images
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Western blot - Anti-eIF3e antibody (ab36766)All lanes : Anti-eIF3e antibody (ab36766) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899)
Lane 3 :A-431 whole cell lysate (ab7909)
Lane 4 :HEK-293 whole cell lysate (ab7902)
Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human eIF3e peptide (ab36902) at 1 µg/ml
Lane 6 :Jurkat whole cell lysate (ab7899) with Human eIF3e peptide (ab36902) at 1 µg/ml
Lane 7 :A-431 whole cell lysate (ab7909) with Human eIF3e peptide (ab36902) at 1 µg/ml
Lane 8 :HEK-293 whole cell lysate (ab7902) with Human eIF3e peptide (ab36902) at 1 µg/ml
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?
Additional bands at: 40 kDa (possible degradation product) -
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 70oC; 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. -
Immunocytochemistry/ Immunofluorescence - Anti-eIF3e antibody (ab36766)ICC/IF image of ab36766 stained human HeLa cells. The cells were methanol fixed (5 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab36766, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue). -
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 BondTM 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.
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Western blot - Anti-eIF3e antibody (ab36766) This image is courtesy of an anonymous AbreviewAll lanes : Anti-eIF3e antibody (ab36766) at 1/1000 dilution
Lane 1 : Chicken tectum lysate
Lane 2 : Zebrafish lysate
Lysates/proteins at 5 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit polyclonal IgG
at 1/10000 dilution
Predicted band size: 52 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?
Additional bands at: 38 kDa (possible degradation product)
