MTOR antibody (AA 2436-2492)

Catalog No. ABIN676403

Product Details anti-MTOR Antibody

Target: MTOR (mTOR) (Mechanistic Target of Rapamycin (serine/threonine Kinase) (mTOR))

Binding Specificity: AA 2436-2492

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This MTOR antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Cross-Reactivity: Chicken, Cow, Fish, Human, Mouse, Rat

Predicted Reactivity: Dog,Cow,Sheep,Horse

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human mTOR

Isotype: IgG

Application Details

Application Notes: IHC-P 1:100-500
IF(IHC-P) 1:50-200

Restrictions: For Research Use only

Independent Validation (WB)

Validation #029761 (Western Blotting)

by: Alamo Laboratories Inc

No.: #029761

Date: 07/03/2014

Lot Number: 131209

Method validated: Western Blotting

Positive Control: HeLa cells

Negative Control: c6/36 mosquito cells (non-reactive species)

Notes: A strong band was observed in the positive control sample at the correct molecular weight. Multiple faint bands were also observed in the positive sample, but these bands did not represent a significant background signal. A band at the correct molecular weight was also observed in the negative control; however, it may be impossible to find a true negative control for this antigen.

Validation Images

Figure 1. Western blot of lysates from HeLa cells (Lane 1) and c6/36 cells (Lane 2) probed with anti-mTOR (upper panel) or with anti-Actin for loading control (lower panel).

Full Methods

Primary Antibody:

• Antigen: Mechanistic Target of Rapamycin (serine/threonine Kinase) (FRAP1) (1:200 dilution)
• Catalog number: ABIN676403
• Lot number: 131209

Secondary Antibody:

• Antigen: Goat Anti-Rabbit IgG (H + L)-HRP Conjugate
• Lot number: L170-6515

Full Protocol:

• Total protein extracts were boiled in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol at 95°C for 5 min prior to loading.
• 20 μg of boiled extracts were loaded and resolved on 8-16% SDS-polyacrylamide gel.
• The Thermo Scientific - Spectra Multicolor Broad Range (Cat # 26634) were used as molecular mass markers.
• Proteins were then transferred onto PVDF membrane by wet transfer and protein transfer was confirmed with Ponceau-S staining.
• The PVDF membrane was incubated with 25 mL of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) BSA at room temperature for 1 h.
• The membrane was rinsed with TBST once.
• The membrane was immersed with the protein side up in the primary antibody solution (FRAP1; 1:200) in TBST containing 5% (W/V) BSA and incubated for 16 h at 4°C.
• The membrane was rinsed in TBST thrice for 5 min each.
• The membrane was incubated in the HRP-conjugated secondary antibody solution (Goat anti-rabbit IgG-HRP; 1:20,000) in TBST containing 5% (W/V) BSA and incubated for 1 h at room temperature (~26°C) with gentle agitation.
• The membrane was rinsed thrice TBST thrice for 5 min each.
• The membrane was rinsed in TBS twice for 30 s each.
• Signals were detected with ECL-2 Substrate. The blot was scanned for 300 s.
• The membrane was rinsed three times TBST.
• Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 min each.
• The membrane was washed in TBST 2 times for 10 min each.
• Repeated Steps 5-12 with the loading control antibody (anti-Actin; 1:6,000) and its matching secondary antibody (Goat anti-rabbit IgG-HRP; 1:20,000).

Experimental Notes: - No experimental challenges noted.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-MTOR antibody (ABIN676403)

Wang, Sun, Han, Li, Ge, Zhao, De Felici, Shen, Cheng: "Protective effects of melatonin against nicotine-induced disorder of mouse early folliculogenesis." in: Aging, Vol. 10, Issue 3, pp. 463-480, (2019) (PubMed).

Sun, Li, Zhao, Wang, Liu, Shao, Xue, Xing: "Synergistic effect of copper and arsenic upon oxidative stress, inflammation and autophagy alterations in brain tissues of Gallus gallus." in: Journal of inorganic biochemistry, Vol. 178, pp. 54-62, (2018) (PubMed).

Li, Qiu, Liu, Wang, Hu, Gan, Wang: "Long-term thermal manipulation in the late incubation period can inhibit breast muscle development by activating endoplasmic reticulum stress in duck (Anasplatyrhynchos domestica)." in: Journal of thermal biology, Vol. 70, Issue Pt B, pp. 37-45, (2017) (PubMed).

Tian, Liu, Qu: "Role of endoplasmic reticulum stress on cisplatin resistance in ovarian carcinoma." in: Oncology letters, Vol. 13, Issue 3, pp. 1437-1443, (2017) (PubMed).

Zou, Geng, Chen, Ren, Liu, Wan, Guo, Wang: "Ranolazine improves oxidative stress and mitochondrial function in the atrium of acetylcholine-CaCl2 induced atrial fibrillation rats." in: Life sciences, Vol. 156, pp. 7-14, (2016) (PubMed).

Madka, Mohammed, Li, Zhang, Biddick, Patlolla, Lightfoot, Towner, Wu, Steele, Kopelovich, Rao: "Targeting mTOR and p53 Signaling Inhibits Muscle Invasive Bladder Cancer In Vivo." in: Cancer prevention research (Philadelphia, Pa.), Vol. 9, Issue 1, pp. 53-62, (2016) (PubMed).

Yang, Wang, Wang, Zhang, Zhang, Lu, Wang: "mTOR is involved in 17?-estradiol-induced, cultured immature boar Sertoli cell proliferation via regulating the expression of SKP2, CCND1, and CCNE1." in: Molecular reproduction and development, Vol. 82, Issue 4, pp. 305-14, (2015) (PubMed).

Li, Liu, Wang, Sun, Ding, Sun, Han, Wang: "Follistatin could promote the proliferation of duck primary myoblasts by activating PI3K/Akt/mTOR signalling." in: Bioscience reports, Vol. 34, Issue 5, (2014) (PubMed).

Target Details for MTOR

Target: MTOR (mTOR) (Mechanistic Target of Rapamycin (serine/threonine Kinase) (mTOR))

Alternative Name: Mtor/Frap (mTOR Products)

Synonyms: FRAP1 antibody, FRAP antibody, FRAP2 antibody, RAFT1 antibody, RAPT1 antibody, 2610315D21Rik antibody, AI327068 antibody, Frap1 antibody, flat antibody, frap1 antibody, tor antibody, wu:fc22h08 antibody, mechanistic target of rapamycin kinase antibody, mechanistic target of rapamycin antibody, mechanistic target of rapamycin (serine/threonine kinase) antibody, MTOR antibody, Mtor antibody, mtor antibody

Background:

Synonyms: Serine/threonine-protein kinase mTOR, FK506-binding protein 12-rapamycin complex-associated protein 1, FKBP12-rapamycin complex-associated protein, Mammalian target of rapamycin, mTOR, Mechanistic target of rapamycin, Rapamycin and FKBP12 target 1, Rapamycin target protein 1, MTOR, FRAP, FRAP1, FRAP2, RAFT1, RAPT1

Background: The protein encoded by this gene belongs to a family of phosphatidylinositol kinase-related kinases. These kinases mediate cellular responses to stresses such as DNA damage and nutrient deprivation. This protein acts as the target for the cell-cycle arrest and immunosuppressive effects of the FKBP12-rapamycin complex. The ANGPTL7 gene is located in an intron of this gene. [provided by RefSeq, Sep 2008]

Gene ID: 2475

UniProt: P42345

Pathways: PI3K-Akt Signaling, RTK Signaling, AMPK Signaling, Interferon-gamma Pathway, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Regulation of Actin Filament Polymerization, Regulation of Muscle Cell Differentiation, Regulation of Cell Size, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Autophagy, CXCR4-mediated Signaling Events, BCR Signaling, Warburg Effect