Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-mTOR antibody [Y391] (ab32028)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y391] to mTOR
  • Suitable for: WB, IP, IHC-P, IHC-Fr
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-mTOR antibody [Y391]
    See all mTOR primary antibodies

  • Description

    Rabbit monoclonal [Y391] to mTOR

  • Host species

    Rabbit

  • Specificity

    Expression levels of the target protein vary with sample type and some optimisation may be required.

  • Tested Applications & Species

    Application Species
    IHC-Fr
    Human
    IHC-P
    Mouse
    Rat
    Human
    IP
    Rat
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human mTOR aa 2400-2500 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab193663)

  • Epitope

    ab32028 reacts with an epitope located in the C terminal region of mTOR.

  • Positive control

    • WB: Jurkat whole cell lysate (ab30128), HeLa, HaCaT and MDA-MB-231 cell lysates and rat brain tissue lysate. IHC-P: Human breast carcinoma, mouse testis and rat testis tissues. IP: Rat brain tissue lysate and HeLa whole cell lysate (ab150035). 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.

Properties

Images

  • Western blot - Anti-mTOR antibody [Y391] (ab32028)
    Western blot - Anti-mTOR antibody [Y391] (ab32028)
    All lanes : Anti-mTOR antibody [Y391] (ab32028) at 1/2000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates in RIPA buffer
    Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates in HOT buffer
    Lane 3 : Rat brain lysates in RIPA buffer
    Lane 4 : Rat brain lysates in HOT buffer
    Lane 5 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
    Lane 6 : Human fetal lung lysates

    Lysates/proteins at 20 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: 290 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Frozen sections) - Anti-mTOR antibody [Y391] (ab32028)
    Immunohistochemistry (Frozen sections) - Anti-mTOR antibody [Y391] (ab32028)

    IHC image of mTOR staining in a section of frozen normal human heart performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab32028, 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.

  • Western blot - Anti-mTOR antibody [Y391] (ab32028)
    Western blot - Anti-mTOR antibody [Y391] (ab32028)
    Lanes 1-7 : Anti-mTOR antibody [Y391] (ab32028)
    Lane 8 : Anti-mTOR antibody [Y391] (ab32028) at 1/1000 dilution

    Lane 1 : Rat brain tissue lysate with 5% NFDM/TBST
    Lane 2 : Rat heart tissue lysate with 5% NFDM/TBST
    Lane 3 : Rat liver tissue lysate with 5% NFDM/TBST
    Lane 4 : Rat spleen tissue with 5% NFDM/TBST
    Lane 5 : Mouse brain tissue lysate with 5% NFDM/TBST
    Lane 6 : Mouse heart tissue lysate with 5% NFDM/TBST
    Lane 7 : Mouse kidney tissue lysate with 5% NFDM/TBST
    Lane 8 : Mouse liver tissue lysate with 5% NFDM /TBST

    Lysates/proteins at 20 µg per lane.

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

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



    Exposure time:

    Lane 1: 10 seconds
    Lane 2-8: 180 seconds

    This antibody detects non-specific bands and high background. It doesn’t detect the target band in some mouse and rat tissues.

  • Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028)
    Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028)

    ab32028 (purified) at a dilution of 1/100 immunoprecipitating mTOR in HeLa whole cell lysate.

    Lane 1 (input): HeLa whole cell lysate (10µg)

    Lane 2 (+): ab32028 + HeLa whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32028 in HeLa whole cell lysate.

    For western blotting, ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028)
    Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028)

    ab32028 (purified) at a dilution of 1/100 immunoprecipitating mTOR in rat brain tissue lysate.

    Lane 1 (input): Rat brain tissue lysate (10µg)

    Lane 2 (+): ab32028 + rat brain tissue lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32028 in rat brain tissue lysate.

    For western blotting, ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Western blot - Anti-mTOR antibody [Y391] (ab32028)
    Western blot - Anti-mTOR antibody [Y391] (ab32028)
    All lanes : Anti-mTOR antibody [Y391] (ab32028) at 1/1000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST
    Lane 2 : HaCaT (Human skin keratinocyte) whole cell lysate with 5% NFDM/TBST
    Lane 3 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST
    Lane 4 : Mouse thymus (8-10 weeks) tissue lysate with 5% NFDM/TBST
    Lane 5 : Mouse lung (8-10 weeks) tissue lysate with 5% NFDM/TBST
    Lane 6 : Rat thymus tissue lysate with 5% NFDM/TBST
    Lane 7 : Rat lung tissue lysate with 5% NFDM/TBST

    Lysates/proteins at 20 µg per lane.

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

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



    Exposure time:

    Lane 1-3: 100 seconds
    Lane 4-7: 180 seconds

    This antibody detects non-specific bands and high background. It doesn’t detect the target band in some mouse and rat tissues.

  • Anti-mTOR antibody [Y391] (ab32028)
    Anti-mTOR antibody [Y391] (ab32028)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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