Recombinant Anti-eIF4EBP1 antibody [Y330] (ab32130)
![Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)](https://www.abcam.com/ps/products/32/ab32130/Images/ab32130-388722-anti-eif4ebp1-antibody-y330-western-blot-hela-lysates.jpg "Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y330] to eIF4EBP1
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-eIF4EBP1 antibody [Y330]
See all eIF4EBP1 primary antibodies -
Description
Rabbit monoclonal [Y330] to eIF4EBP1 -
Host species
Rabbit -
Specificity
ab32130 recognises 4E-BP1 (eIF4E-binding protein) also known as PHAS. -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HEK-293 cell lysate, HAP1 and K562 cell lysate, IHC-P: Human prostate carcinoma, Flow Cyt (intra): HeLa cells
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y330 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32130 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 |
1/1000. Detects a band of approximately 15-20 kDa (predicted molecular weight: 12 kDa).
|
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| IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| Flow Cyt (Intra) |
1/1000.
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| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 15-20 kDa (predicted molecular weight: 12 kDa). |
|
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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Flow Cyt (Intra)
1/1000. |
Target
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Function
Regulates eIF4E activity by preventing its assembly into the eIF4F complex. Mediates the regulation of protein translation by hormones, growth factors and other stimuli that signal through the MAP kinase and mTORC1 pathways. -
Sequence similarities
Belongs to the eIF4E-binding protein family. -
Post-translational
modificationsPhosphorylated on serine and threonine residues in response to insulin, EGF and PDGF. Phosphorylation at Thr-37, Thr-46, Ser-65 and Thr-70 is regulated by mTORC1. Phosphorylated upon DNA damage, probably by ATM or ATR. - Information by UniProt
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Database links
- Entrez Gene: 1978 Human
- Omim: 602223 Human
- SwissProt: Q13541 Human
- Unigene: 411641 Human
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Alternative names
- 4E-BP1 antibody
- 4EBP1 antibody
- 4EBP1_HUMAN antibody
see all
Images
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Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)All lanes : Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : EIF4EBP1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 12 kDa
Observed band size: 13 kDa why is the actual band size different from the predicted?Lanes 1-2: Merged signal (red and green). Green - ab32130 observed at 13 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32130 Anti-eIF4EBP1 antibody [Y330] was shown to specifically react with eIF4EBP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264784 (knockout cell lysate ab257146) was used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32130 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y330] (ab32130)](https://www.abcam.com/ps/products/32/ab32130/Images/ab32130-43977-anti-eif4ebp1-antibody-y330-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y330] (ab32130)ab32130, staining human prostate carcinoma by immunohistochemistry, Paraffin embedded tissue
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Flow Cytometry (Intracellular) - Anti-eIF4EBP1 antibody [Y330] (ab32130)](https://www.abcam.com/ps/products/32/ab32130/Images/ab32130-6-anti-eif4ebp1-antibody-y330-flowcytometry-hela-human.jpg)
Flow Cytometry (Intracellular) - Anti-eIF4EBP1 antibody [Y330] (ab32130)Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling eIF4EBP1 with purified ab32130 at 1/1000 dilution (1 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Left).
Low expression in mitotic phase was consistent with literature. (PMID: 11553333) -
![Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)](https://www.abcam.com/ps/products/32/ab32130/Images/ab32130-256326-anti-eif4ebp1-antibody-y330-western-blot.jpg)
Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)All lanes : Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : eIF4EBP1 knockout HAP1 cell lysate
Lane 3 : Hek293 cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 12 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32130 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32130 was shown to specifically react with eIF4EBP1 when eIF4EBP1 knockout samples were used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32130 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
![Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)](https://www.abcam.com/ps/products/32/ab32130/Images/ab32130-43978-anti-eif4ebp1-antibody-y330-western-blot.jpg)
Western blot - Anti-eIF4EBP1 antibody [Y330] (ab32130)Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution + K562 cell lysate
Predicted band size: 12 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted? -
![Anti-eIF4EBP1 antibody [Y330] (ab32130)](https://www.abcam.com/ps/products/32/ab32130/Images/ab32130-5-benefits-of-recombinant-antibodies.png)
Anti-eIF4EBP1 antibody [Y330] (ab32130)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab32130 has been referenced in 9 publications.
- Liu L et al. Se improves GPX4 expression and SOD activity to alleviate heat-stress-induced ferroptosis-like death in goat mammary epithelial cells. Anim Cells Syst (Seoul) 25:283-295 (2021). PubMed: 34745435
- Guo Y et al. Spontaneous formation of neutrophil extracellular traps is associated with autophagy. Sci Rep 11:24005 (2021). PubMed: 34907287
- Batool A et al. Eukaryotic Initiation Factor 4E phosphorylation acts a switch for its binding to 4E-BP1 and mRNA cap assembly. Biochem Biophys Res Commun 527:489-495 (2020). PubMed: 32336547
- Hori Y et al. Analysis of mTOR pathway expression in lymphatic malformation and related diseases. Pathol Int 70:323-329 (2020). PubMed: 32067331
- Wu X et al. PI3K/AKT/mTOR pathway inhibitors inhibit the growth of melanoma cells with mTOR H2189Y mutations in vitro. Cancer Biol Ther 19:584-589 (2018). PubMed: 29708815
- Wang W et al. Rasfonin promotes autophagy and apoptosis via upregulation of reactive oxygen species (ROS)/JNK pathway. Mycology 7:64-73 (2016). PubMed: 30123617
- Lu Q et al. Akt inhibition attenuates rasfonin-induced autophagy and apoptosis through the glycolytic pathway in renal cancer cells. Cell Death Dis 6:e2005 (2015). Human . PubMed: 26633711
- Hu Y et al. MiR-99a antitumor activity in human breast cancer cells through targeting of mTOR expression. PLoS One 9:e92099 (2014). WB ; Human . PubMed: 24637915
- Fukuda A et al. Growth-dependent podocyte failure causes glomerulosclerosis. J Am Soc Nephrol 23:1351-63 (2012). PubMed: 22773827