WB result of FTO Recombinant Rabbit mAb
Primary antibody: FTO Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: SH-SY5Y whole cell lysate 20 µg
Lane 3: 293T whole cell lysate 20 µg
Lane 4: U-87 MG whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 58 kDa
Observed MW: 58 kDa
FTO Recombinant Rabbit mAb (S-1663-48)
FTO Recombinant Rabbit mAb (S-1663-48)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | FTO |
| Synonyms | Alpha-ketoglutarate-dependent dioxygenase FTO; Fat mass and obesity-associated protein; U6 small nuclear RNA (2'-O-methyladenosine-N(6)-)-demethylase FTO; U6 small nuclear RNA N(6)-methyladenosine-demethylase FTO; mRNA (2'-O-methyladenosine-N(6)-)-demethylase FTO (m6A(m)-demethylase FTO); mRNA N(6)-methyladenosine demethylase FTO; tRNA N1-methyl adenine demethylase FTO; KIAA1752 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Nucleus |
| Accession | Q9C0B1 |
| Clone Number | S-1663-48 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, IP |
| Reactivity | Hu, Ms, Rt |
| Positive Sample | HeLa, SH-SY5Y, 293T, U-87 MG, NIH/3T3, C2C12, PC-12 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms, Rt |
| IP | 1:50 | Hu |
| IHC-P | 1:200 | Hu, Ms, Rt |
| ICC | 1:500 | Hu, Ms |
Background
FTO (Fat mass and obesity-associated protein) is an enzyme that acts as an RNA N6-methyladenosine (m6A) demethylase. FTO is the first identified demethylase for internal m6A modification in RNA, which is a dynamic and reversible process. It has the capability to remove the methyl group from m6A and N6,2-O-dimethyladenosine (m6Am) in RNA, primarily found in the 5’-UTR. FTO has been reported to be upregulated in many tumors, and its high expression is associated with shorter overall survival in multiple types of cancer patients. It plays an oncogenic role in acute myeloid leukemia (AML) and is involved in cancer stem cell self-renewal and immune evasion. Targeting FTO suppresses cancer stem cell maintenance and immune evasion, suggesting its critical roles in these processes and highlighting the potential of targeting FTO for cancer therapy. FTO is also implicated in epitranscriptomics, which involves chemical modifications in RNA that affect the fate of modified RNAs.
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Western Blot
WB result of FTO Recombinant Rabbit mAb
Primary antibody: FTO Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Lane 2: C2C12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 58 kDa
Observed MW: 58 kDa
WB result of FTO Recombinant Rabbit mAb
Primary antibody: FTO Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: PC-12 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 58 kDa
Observed MW: 58 kDa
IP
FTO Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating FTO in 0.4 mg SH-SY5Y whole cell lysate.
Western blot was performed on the immunoprecipitate using FTO Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: SH-SY5Y whole cell lysate 20 µg (Input)
Lane 2: FTO Rabbit mAb IP in SH-SY5Y whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in SH-SY5Y whole cell lysate
Predicted MW: 58 kDa
Observed MW: 58 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human kidney. Anti-FTO antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human breast cancer. Anti-FTO antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human clear cell carcinoma of kidney. Anti-FTO antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse kidney. Anti-FTO antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat kidney. Anti-FTO antibody was used at 1/200 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in SH-SY5Y cells. Anti-FTO antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ICC shows positive staining in NIH/3T3 cells. Anti-FTO antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
