Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP2151Y] to eIF4E (phospho S209)
Suitable for: ICC/IF, WB, IP, IHC-P, Dot blot
Reacts with: Mouse, Rat, Human, Pig

Product name

Anti-eIF4E (phospho S209) antibody [EP2151Y]
See all eIF4E primary antibodies
Description

Rabbit monoclonal [EP2151Y] to eIF4E (phospho S209)
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Mouse Human
IHC-P	Human
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide within Human eIF4E. The exact sequence is proprietary.
Database link: P06730
Positive control
- WB: 293 cell lysate treated with alkaline phosphatase and HEK293 cell lysate treated with Dexamethasone. IHC-P: human breast carcinoma tissue. ICC/IF: HEK293 cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP2151Y
Isotype

IgG
Research areas

Western blot - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/100000 dilution (purified)

Lane 1 : Untreated HEK293 whole cell lysate
Lane 2 : HEK293 cells treated with 10uM dexamethasone for 1 hour whole cell lysate
Lane 3 : HEK293 cells treated with 10uM dexamethasone for 1 hour whole cell lysate.The membrane was then incubated with alkaline phosphatase.

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 30 seconds

Blocking buffer and concentration 2% BSA/TBST.
Diluting buffer and concentration 2% BSA/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling eIF4E with purified ab76256 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

Immunocytochemistry/Immunofluorescence analysis of untreated, 20% serum treated and 20% serum + LP treated NIH/3T3 cells labelling eIF4E (phospho S209) with ab76256 (left) and eIF4E with ab33766 (right) both at a dilution of 1/500.

Cells were fixed with 100% methanol. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

The image shows increased cytoplasmic staining after 20% serum treatment on NIH3T3 cells when compared with no serum treated cells. The LP treatment decreased the increased cytoplasmic staining caused by 20% serum.

ab33766 was used as a Pan control for ab76256. The results showed cytoplasmic staining on no serum, 20% serum and 20% serum +LP treated NIH3T3 cells.
Immunoprecipitation - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

ab76256 (purified) at 1/40 immunoprecipitating eIF4E (phospho S209) in HEK293 whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/1,500) was used for detection. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/1000 dilution (purified)

Lane 1 : Mouse spleen lysate
Lane 2 : Rat brain lysate
Lane 3 : Pig heart lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 25 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Dot Blot - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

Dot blot analysis of eIF4E (pS209) peptide (Lane 1) and eIF4E non-phospho peptide (Lane 2) labelling eIF4E (pS209) with purified ab76256 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Immunocytochemistry/ Immunofluorescence - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

Immunocytochemistry/Immunofluorescence analysis of serum starved HEK293 cells treated with CGP 57380 ab120365) labelling eIF4E (phospho S209) with unpurified ab32124 at 1/100. Decrease in eIF4E (phospho S209) expression correlates with increased concentration of CGP 57380, as described in literature.
The cells were incubated at 37°C for 1h in media containing different concentrations of ab120365 (CGP 57380) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with unpurified ab76256 was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Western blot - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/50000 dilution (purified)

Lane 1 : Untreated HEK293 cell lysate
Lane 2 : HEK293 treated with 10mM Dexamethasone 1 hour lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 25 kDa

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/50000 dilution (purified)

Lane 1 : Untreated 293 cell lysate
Lane 2 : 293 cell lysate treated with alkaline phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 25 kDa

Exposure time: 1 minute

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

All lanes : Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256) at 1/100000 dilution (purified)

Lane 1 : Untreated HEK293 cell lysate
Lane 2 : HEK293 cell lysate - treated with Dexamethasone

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 25 kDa

Exposure time:
eIF4E pS209: 15 seconds.
eIF4E: 3 minutes.

Blocking and dilution buffer: 5% NFDM/TBST.
Anti-eIF4E (phospho S209) antibody [EP2151Y] (ab76256)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com