Anti-eIF3B antibody (ab40799)
Key features and details
- Rabbit polyclonal to eIF3B
- Suitable for: WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-eIF3B antibody
See all eIF3B primary antibodies -
Description
Rabbit polyclonal to eIF3B -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Cow, Drosophila melanogaster
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 - 250 of Human eIF3B.
Read Abcam's proprietary immunogen policy (Peptide available as ab41603.) -
Positive control
- This antibody gave a positive signal in the following whole cell lysates; HEK293, HepG2, SHSY-5Y and U2OS. IF/ICC: HeLa cell line.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab40799 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 92 kDa). ICC/IF Use a concentration of 5 µg/ml. Target
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Function
Component of the eukaryotic translation initiation factor 3 (eIF-3) complex, which is required for several steps in the initiation of protein synthesis. The eIF-3 complex associates with the 40S ribosome and facilitates the recruitment of eIF-1, eIF-1A, eIF-2:GTP:methionyl-tRNAi and eIF-5 to form the 43S preinitiation complex (43S PIC). The eIF-3 complex stimulates mRNA recruitment to the 43S PIC and scanning of the mRNA for AUG recognition. The eIF-3 complex is also required for disassembly and recycling of post-termination ribosomal complexes and subsequently prevents premature joining of the 40S and 60S ribosomal subunits prior to initiation. -
Sequence similarities
Belongs to the eIF-3 subunit B family.
Contains 1 RRM (RNA recognition motif) domain.
Contains 5 WD repeats. -
Domain
The RRM domain mediates interaction with EIF3J. -
Post-translational
modificationsPhosphorylated. Phosphorylation is enhanced upon serum stimulation. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 789999 Cow
- Entrez Gene: 36981 Drosophila melanogaster
- Entrez Gene: 8662 Human
- Entrez Gene: 27979 Mouse
- Entrez Gene: 288516 Rat
- Omim: 603917 Human
- SwissProt: A7MB16 Cow
- SwissProt: Q0E940 Drosophila melanogaster
see all -
Alternative names
- eIF-3-eta antibody
- eIF3 eta antibody
- eIF3 p110 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-eIF3B antibody (ab40799)
ICC/IF image of ab40799 stained MCF7 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 (ab40799, 5µg/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 1.43µM.
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Western blot - Anti-eIF3B antibody (ab40799)All lanes : Anti-eIF3B antibody (ab40799) at 1 µg/ml
Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 4 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
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: 92 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
This antibody was raised against an immunogen that is predicted to cross react with both isoforms 1 and 2 of eIF3B. We believe that the band observed is isoform 2 due to its higher molecular weight of 110 kDa. Predicted molecular weights of the isoforms are 92 kDa and 99 kDa respectively.
Protocols
References (4)
ab40799 has been referenced in 4 publications.
- Wang L & Ouyang L Effects of EIF3B gene downregulation on apoptosis and proliferation of human ovarian cancer SKOV3 and HO-8910 cells. Biomed Pharmacother 109:831-837 (2019). PubMed: 30551537
- Tang L et al. Muscleblind-like 1 destabilizes Snail mRNA and suppresses the metastasis of colorectal cancer cells via the Snail/E-cadherin axis. Int J Oncol 54:955-965 (2019). PubMed: 30664186
- Hodgson RE et al. Cellular eIF2B subunit localization: implications for the integrated stress response and its control by small molecule drugs. Mol Biol Cell 30:942-958 (2019). PubMed: 30726166
- Fathinajafabadi A et al. CERKL, a retinal disease gene, encodes an mRNA-binding protein that localizes in compact and untranslated mRNPs associated with microtubules. PLoS One 9:e87898 (2014). WB ; Human . PubMed: 24498393
Images
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Immunocytochemistry/ Immunofluorescence - Anti-eIF3B antibody (ab40799)
ICC/IF image of ab40799 stained MCF7 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 (ab40799, 5µg/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 1.43µM.
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Western blot - Anti-eIF3B antibody (ab40799)All lanes : Anti-eIF3B antibody (ab40799) at 1 µg/ml
Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 4 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
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: 92 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
This antibody was raised against an immunogen that is predicted to cross react with both isoforms 1 and 2 of eIF3B. We believe that the band observed is isoform 2 due to its higher molecular weight of 110 kDa. Predicted molecular weights of the isoforms are 92 kDa and 99 kDa respectively.
