Human EIF4EBP1 knockout HeLa cell lysate
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EIF4EBP1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
View Alternative Names
4EBP1_HUMAN, BP 1, Eif4ebp1, Eukaryotic translation initiation factor 4E-binding protein 1, PHAS-I, Phosphorylated heat- and acid-stable protein regulated by insulin 1, eIF4E-binding protein 1
- WB
Lab
Western blot - Human EIF4EBP1 knockout HeLa cell lysate (AB257146)
Lane 1 : Wild-type HeLa cell lysate (20 μg)
Lane 2 : EIF4EBP1 knockout HeLa cell lysate (20 μg)
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.
All lanes:
Western blot - Anti-eIF4EBP1 antibody [Y330] (<a href='/en-us/products/primary-antibodies/eif4ebp1-antibody-y330-ab32130'>ab32130</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
EIF4EBP1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human EIF4EBP1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-eif4ebp1-knockout-hela-cell-line-ab264784'>ab264784</a>)
Predicted band size: 13 kDa,36 kDa
Observed band size: 13 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human EIF4EBP1 knockout HeLa cell lysate (AB257146)
Homozygous : Insertion of the selection cassette in exon 1
- WB
Lab
Western blot - Human EIF4EBP1 knockout HeLa cell lysate (AB257146)
Lane 1 : Wild-type HeLa cell lysate (20 µg)
Lane 2 : EIF4EBP1 knockout HeLa cell lysate (20 µg)
Lanes 1-2 : Merged signal (red and green). Green - ab32024 observed at 13 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32024 Anti-eIF4EBP1 antibody [Y329] 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. ab32024 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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.
All lanes:
Western blot - Anti-eIF4EBP1 antibody [Y329] (<a href='/en-us/products/primary-antibodies/eif4ebp1-antibody-y329-ab32024'>ab32024</a>) at 1/5000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human EIF4EBP1 knockout HeLa cell lysate (ab257146) at 20 µg
Secondary
Lanes 1 - 2:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Lanes 1 - 3:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 13 kDa
Observed band size: 13 kDa,37 kDa
false
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
EIF4EBP1 plays an important role in regulating cell growth and proliferation by modulating protein synthesis. It is a part of the eIF4F complex which is responsible for the initiation of mRNA translation. When hypophosphorylated eIF4EBP1 binds tightly to eIF4E and prevents the assembly of the active eIF4F complex leading to reduced translation initiation of mRNAs involved in growth and survival. This regulation is important in conditions where cells need to adapt to metabolic stress or external signals.
Pathways
The mammalian target of rapamycin (mTOR) pathway regulates eIF4EBP1 through phosphorylation. In response to growth signals mTOR phosphorylates eIF4EBP1 causing the release of eIF4E and allowing mRNA translation to proceed. This interaction links eIF4EBP1 to the PI3K/AKT/mTOR signaling pathway which influences cell cycle progression and survival. Related proteins in this pathway include ribosomal protein S6 kinase (S6K1) which is also phosphorylated by mTOR to promote protein synthesis.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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