FTO Monoclonal antibody

FTO Monoclonal Antibody for WB, IHC, ELISA, FC (Intra)

Host / Isotype

Mouse / IgG2b

Reactivity

Human, Mouse, Pig and More (3)

Applications

WB, IHC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

3

CloneNo.

1E8B1

Cat no : 68111-1-Ig

Synonyms

KIAA1752, 1E8B1

---

Validation Data Gallery

• 

  WB analysis using 68111-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68111-1-Ig (FTO antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

• 

  WB analysis of NIH/3T3 using 68111-1-Ig

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 68111-1-Ig (FTO antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of pig brain using 68111-1-Ig

  pig brain tissue were subjected to SDS PAGE followed by western blot with 68111-1-Ig (FTO antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of mouse brain using 68111-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 68111-1-Ig (FTO antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 68111-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 68111-1-Ig (FTO antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human liver cancer using 68111-1-Ig

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 68111-1-Ig (FTO antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 68111-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 68111-1-Ig (FTO antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  FC experiment of SH-SY5Y using 68111-1-Ig

  1X10^6 SH-SY5Y cells were intracellularly stained with 0.4 ug Anti-Human FTO (68111-1-Ig, Clone:1E8B1) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2b Isotype Control (MPC-11) (65128-1-Ig, Clone: MPC-11) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

---

Tested Applications

• Positive WB detected in: LNCaP cells, NIH/3T3 cells, pig brain tissue, HeLa cells, HEK-293 cells, Jurkat cells, K-562 cells
• Positive IHC detected in: mouse brain tissue, human liver cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive FC (Intra) detected in: SH-SY5Y cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 3 publications below

Product Information

68111-1-Ig targets FTO in WB, IHC, FC (Intra), ELISA applications and shows reactivity with Human, Mouse, Pig samples.

• Tested Reactivity: Human, Mouse, Pig
• Cited Reactivity: human, mouse, pig
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: FTO fusion protein Ag26095
• Full Name: fat mass and obesity associated
• Calculated Molecular Weight: 58 kDa
• Observed Molecular Weight: 58 kDa
• GenBank Accession Number: NM_001080432
• Gene Symbol: FTO
• Gene ID (NCBI): 79068
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Fat mass and obesity-associated protein (FTO) has efficient oxidative demethylation activity targeting the abundant N6-methyladenosine (m6A) residues in RNA in vitro. Variants in the FTO (fat mass and obesity associated) gene are associated with increased body mass index in humans.

Publications

Species	Application	Title
pig	WB	Int J Biol Macromol; METTL3 enhances E. coli F18 resistance by targeting IKBKG/NF-κB signaling via an m6A-YTHDF1-dependent manner in IPEC-J2 cells; Authors - Jian Jin
mouse	WB	Acta Physiol (Oxf); Landscape of transcriptome-wide m6A modification in diabetic liver reveals rewiring of PI3K-Akt signaling after physical exercise; Authors - Cong Chen
human	WB	Apoptosis; NRP1 contributes to stemness and potentiates radioresistance via WTAP-mediated m6A methylation of Bcl-2 mRNA in breast cancer; Authors - Yang Wang