Antibodies, Proteins, Kits and Reagents for Life Science

365 193 ₸ Product size

100 µl 365 193 ₸ 10 µl 144 067 ₸

Order now and get it on Tuesday March 02, 2021

Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR426(2)] to mTOR (phospho S2448)
Suitable for: WB, IHC-P, Dot blot, IHC-Fr
Reacts with: Mouse, Human

Product name

Anti-mTOR (phospho S2448) antibody [EPR426(2)]
See all mTOR primary antibodies
Description

Rabbit monoclonal [EPR426(2)] to mTOR (phospho S2448)
Host species

Rabbit
Specificity

This antibody only detects mTOR/FRAP phosphorylated at serine 2448.

Tested Applications & Species

Application	Species
IHC-Fr	Human
IHC-P	Human
WB	Mouse Human

See all applications and species data

Immunogen

This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK293, HeLa and NIH/3T3 cell lysates. IHC-P: Human breast and endometrium carcinoma tissue. Dot Blot: mTOR (phospho S2448) phospho peptide. IHC-Fr: Human heart tissue sections.
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.

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: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR426(2)
Isotype

IgG
Research areas

Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/2000 dilution (Purified)

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa grown in serum-free media overnight, then treated with 200nM PMA for 4 hours whole cell lysate
Lane 3 : HeLa grown in serum-free media overnight, then treated with 200nM PMA for 4 hours whole cell lysate. Then the membrane was 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: 289 kDa
Observed band size: 289 kDa

Blocking buffer: 5% NFDM/TBST
Dilutiing buffer: 5% NFDM /TBST
Immunohistochemistry (Frozen sections) - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

IHC image of mTOR (phospho S2448) staining in a section of frozen normal human heart performed on a Leica BOND^TM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab109268, 1/200 dilution, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using unpurified ab109268 at a dilution of 1/50. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Dot Blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

Dot blot analysis of mTOR (phospho S2448) phospho peptide (Lane 1) and mTOR non-phospho peptide (Lane 2) labeling mTOR (phospho S2448) phospho peptide with purified ab109268 at a dilution of 1/1000 (0.073ug/ml). A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100,000.

Blocking buffer: 5% NFDM/TBST
Diluting buffer: 5% NFDM /TBST
Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/1000 dilution (purified)

Lane 1 : untreated NIH/3T3 cell lysate
Lane 2 : NIH/3T3 cell lysate treated with insulin
Lane 3 : NIH/3T3 cell lysate treated with insulin, then the membrane treated wth alkaline phosphatase

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 289 kDa
Observed band size: 289 kDa

Exposure time: 2 minutes

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

Immunohistochemical cytoplasmic and nuclear staining of paraffin embedded human endometrium carcinoma with purified ab109268 at a working dilution of 1 in 100. The secondary antibody used is ab97051, a HRP goat anti-rabbit IgG (H+L), at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/1000 dilution (purified)

Lane 1 : untreated HeLa cell lysate
Lane 2 : HeLa cell lysate treated with insulin
Lane 3 : HeLa cell lystae treated with insulin, and the membrane treated with alkaline phosphatase

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution (HRP goat anti-rabbit (H+L))

Predicted band size: 289 kDa
Observed band size: 289 kDa

Exposure time: 1 minute

Blocking buffer: 2% BSA/TBST
Dilution buffer: 2% BSA/TBST
Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/2000 dilution (purified)

Lane 1 : untreated HEK293 cell lysate
Lane 2 : HEK293 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 (HRP goat anti-rabbit IgG (H+L))

Predicted band size: 289 kDa
Observed band size: 289 kDa

Exposure time: 3 minutes

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/5000 dilution (unpurified)

Lane 1 : untreated HEK293 cell lysate
Lane 2 : HEK293 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 1/2000 dilution (HRP goat anti-rabbit IgG (H+L))

Predicted band size: 289 kDa
Observed band size: 289 kDa

Exposure time: 3 minutes

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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