Immunocytochemistry (ICC) analysis of HeLa Cl6 cells with anti-mTOR antibody (Cat. No. 490947). HeLa cells were blocked with 5% BSA in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-mTOR rabbit polyclonal was diluted in blocking buffer (antibody concentration 1 ug/ml) and incubated over-night at 4°C. The secondary antibody goat anti-rabbit Alexa Fluor 488 conjugate was also diluted in blocking buffer (antibody concentration 2.5 ug/ml) and incubated for 30 min at 37°C and 30 min at RT. DNA was stained with DAPI in blue.
Western blot analysis of HEK lysate with anti-mTOR antibody (Cat. No. 490947). Western blot analysis was performed on 20 ug wild type (WT) HEK lysate and 20 ug mTOR knock-out (KO) HEK lysate. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-mTOR rabbit polyclonal was diluted in blocking buffer (antibody concentration 500 ng/ml) and incubated for 1 h at RT. The secondary antibody goat anti-rabbit IgG HRP (Cat. No. 90003) was also diluted in blocking buffer (antibody concentration 200 ng/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Immunohistochemistry (IHC) analysis of human head and neck squamous cell carcinoma using anti-mTOR antibody (Cat. No. 490947). IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-mTOR, rabbit polyclonal was diluted in PBS (antibody concentration 10 ug/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visible and with Haemalaun for a few minutes. The 20x picture was acquired using microscopy (courtesy of J.Hess, University Hospital Heidelberg).
Western blot analysis of HEK and HeLa lysate with anti-mTOR antibody (Cat. No. 490947). Western blot analysis was performed on 50 ug HeLa lysate and 40 ug HEK lysate. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1.5 h at RT. The primary antibody anti-mTOR rabbit polyclonal was diluted in blocking buffer (antibody concentration 250 ng/ml) and incubated over-night at 4°C. The secondary antibody goat anti-rabbit IgG HRP (Cat. No. 90003) was also diluted in blocking buffer (antibody concentration 200 ng/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Immunocytochemistry (ICC) analysis of HeLa Cl6 cells with anti-mTOR antibody (Cat. No. 490947). HeLa cells were blocked with 5% BSA in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-mTOR rabbit polyclonal was diluted in blocking buffer (antibody concentration 1 ug/ml) and incubated over-night at 4°C. The secondary antibody goat anti-rabbit Alexa Fluor 488 conjugate was also diluted in blocking buffer (antibody concentration 2.5 ug/ml) and incubated for 30 min at 37°C and 30 min at RT. DNA was stained with DAPI in blue.
Western blot analysis of HEK lysate with anti-mTOR antibody (Cat. No. 490947). Western blot analysis was performed on 20 ug wild type (WT) HEK lysate and 20 ug mTOR knock-out (KO) HEK lysate. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1 h at RT. The primary antibody anti-mTOR rabbit polyclonal was diluted in blocking buffer (antibody concentration 500 ng/ml) and incubated for 1 h at RT. The secondary antibody goat anti-rabbit IgG HRP (Cat. No. 90003) was also diluted in blocking buffer (antibody concentration 200 ng/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Immunohistochemistry (IHC) analysis of human head and neck squamous cell carcinoma using anti-mTOR antibody (Cat. No. 490947). IHC was performed on formalin fixed paraffin embedded sections. The samples were deparaffinized with xylol and ethanol followed by heat induced antigen retrieval with 10 mM citrate buffer. After preparation the tissue was blocked with normal serum for 20 min at RT. The primary antibody anti-mTOR, rabbit polyclonal was diluted in PBS (antibody concentration 10 ug/ml) and incubated at 4°C over-night. The secondary antibody ImmPRESS HRP anti-mouse IgG was incubated for 20 min at RT. Slides were incubated with DAB solution until a brown staining is visible and with Haemalaun for a few minutes. The 20x picture was acquired using microscopy (courtesy of J.Hess, University Hospital Heidelberg).
Western blot analysis of HEK and HeLa lysate with anti-mTOR antibody (Cat. No. 490947). Western blot analysis was performed on 50 ug HeLa lysate and 40 ug HEK lysate. The PVDF membrane was blocked with 5% dry milk in PBST (PBS + 0.1% Tween 20) for 1.5 h at RT. The primary antibody anti-mTOR rabbit polyclonal was diluted in blocking buffer (antibody concentration 250 ng/ml) and incubated over-night at 4°C. The secondary antibody goat anti-rabbit IgG HRP (Cat. No. 90003) was also diluted in blocking buffer (antibody concentration 200 ng/ml) and incubated for 1 h at RT. The bands were visualized by chemiluminescent detection using PierceTM ECL Western Blotting Substrate.
Key Features
- Rabbit polyclonal
- Suitable for IHC, ICC/IF, IP and WB
- Reacts with human, mouse, dog, monkey and rat
- Knock out validated
Product description
| Quantity | 100 µl |
|---|---|
| Antibody Type | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Application | ICC/IF, IHC, IP, WB |
| Purification | Affinity chromatography |
| Reactivity | Dog, Human, Monkey, Mouse, Rat |
| Storage | Up to 3 months at 2-8°C; long term storage in aliquots at -20°C; avoid freeze/thaw cycles |
| Intended use | Research use only |
| Immunogen | Peptide from human mTOR (GTTVPESIHSFIGDGLVKPE) |
| Concentration | 500 µg/ml |
| Formulation | PBS, pH 7.4 with 0.09% sodium azide and 0.5% BSA |
| UniprotID | P42345 (Human) |
| Synonym | Mechanistic target of rapamycin, Mammalian target of rapamycin |
| Note | Centrifuge prior to opening |
Applications
| Tested applications | Tested dilutions |
|---|---|
| Immunocytochemistry (ICC)/ Immunofluorescence (IF) | 1:500-1:2,000 |
| Immunohistochemistry (IHC) - paraffin | 1:50-1:100 (microwave treatment recommended) |
| Immunoprecipitation (IP) | 10 µg |
| Western Blot (WB) | 1:500-1:2,000 |
Background
Mammalian target of rapamycin (mTOR) is a serine/threonine kinase that plays a key role in protein synthesis, cell growth, proliferation and differentiation. It is inhibited by rapamycin, which has immunosuppressant functions. The 290 kDa protein is highly regulated by growth factors and nutrients like glucose and amino acids. mTOR is involved in several pathways from protein degradation via autophagy to protein synthesis via translation initiation (4EBP), ribosome and lipid synthesis. It is located in vesicles and the cytosol.
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FAQs
Most of our purified mouse antibodies contain 0.5% BSA as stabilizer. If BSA was added to the antibody solution, it is stated in the datasheet.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
The supernatant format contains FCS proteins from cell culture medium supplemented with FCS.
The serum antibodies contain other proteins present in serum.
Lyophilized antibodies can be stored at 2-8°C until expiration.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
Most of our liquid antibodies and reconstituted lyophilized antibodies may be stored for short term storage (up to 3 month) at 2-8°C. For long term storage we recommend to store the antibody at -20°C in aliquots. Please avoid freeze and thaw cycles.
Most of our conjugated antibodies should be stored at 2-8°C.
The individual storage conditions are mentioned on the datasheet.
The expiration date of our antibodies is indicated on the product label.
Most of our antibodies contain 0.09% sodium azide as preservative. If a preservative is added, it is mentioned in the datasheet.
The optimal antibody dilution for your specific protocol and application needs to be titrated in your lab with your equipment and sample. The optimal dilution may vary between protocols and samples. A good dilution for starting the titration is the dilution mentioned in the datasheet. If the sample needs a specific treatment (eg. Antigen retrieval for IHC on FFPE sections) this should also be mentioned on the antibody datasheet.
PROGEN antibodies are shipped at ambient temperature. The antibodies are stable at ambient temperature for the shipment period. Please store the antibodies as indicated in the datasheet upon arrival.
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