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  1. Link

    fto-antibody-ab94482.pdf

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Neuroscience Neurology process Metabolism
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Validated using a knockout cell line

Anti-FTO antibody (ab94482)

  • Datasheet
Submit a review Q&A (1)References (4)

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Western blot - Anti-FTO antibody (ab94482)
  • Western blot - Anti-FTO antibody (ab94482)

Key features and details

  • Rabbit polyclonal to FTO
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-FTO antibody
    See all FTO primary antibodies
  • Description

    Rabbit polyclonal to FTO
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Sheep, Rabbit, Cow, Dog, Pig, Orangutan
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • This antibody gave a positive signal in HepG2 whole cell lysate as well as the following tissue lysates: Human brain; Mouse hippocampus; Mouse brain.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Metabolism
    • Cardiovascular
    • Atherosclerosis
    • Lipoprotein metabolism
    • Cardiovascular
    • Atherosclerosis
    • Lipid transport
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipoprotein metabolism
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Obesity
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease
    • Metabolism
    • Types of disease
    • Metabolic disorders

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human FTO protein (ab109039)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab94482 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).

Target

  • Function

    Dioxygenase that repairs alkylated DNA and RNA by oxidative demethylation. Has highest activity towards single-stranded RNA containing 3-methyluracil, followed by single-stranded DNA containing 3-methylthymine. Has low demethylase activity towards single-stranded DNA containing 1-methyladenine or 3-methylcytosine. Has no activity towards 1-methylguanine. Has no detectable activity towards double-stranded DNA. Requires molecular oxygen, alpha-ketoglutarate and iron. Contributes to the regulation of the global metabolic rate, energy expenditure and energy homeostasis. Contributes to the regulation of body size and body fat accumulation.
  • Tissue specificity

    Ubiquitously expressed, with relatively high expression in adrenal glands and brain; especially in hypothalamus and pituitary.
  • Involvement in disease

    Defects in FTO are the cause of growth retardation developmental delay coarse facies and early death (GRDDCFED) [MIM:612938]. The disease consists of a severe children multiple congenital anomaly syndrome with death by the age of 3 years. All affected individuals had postnatal growth retardation, microcephaly, severe psychomotor delay, functional brain deficits, and characteristic facial dysmorphism. In some patients, structural brain malformations, cardiac defects, genital anomalies, and cleft palate were also observed.
  • Sequence similarities

    Belongs to the fto family.
  • Domain

    The 3D-structure of the Fe2OG dioxygenase domain is similar to that of the Fe2OG dioxygenase domain found in the bacterial DNA repair dioxygenase alkB and its mammalian orthologs, but sequence similarity is very low. As a consequence, the domain is not detected by protein signature databases.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession Q9C0B1 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 79068 Human
    • Entrez Gene: 26383 Mouse
    • Entrez Gene: 291905 Rat
    • Omim: 610966 Human
    • SwissProt: Q9C0B1 Human
    • SwissProt: Q8BGW1 Mouse
    • SwissProt: Q2A121 Rat
    • Unigene: 528833 Human
    • Unigene: 4375 Mouse
    • Unigene: 11858 Rat
    see all
  • Alternative names

    • AlkB homolog 9 antibody
    • ALKBH9 antibody
    • Alpha-ketoglutarate-dependent dioxygenase FTO antibody
    • AW743446 antibody
    • Fat mass and obesity-associated protein antibody
    • FATSO, MOUSE, HOMOLOG OF antibody
    • Fto antibody
    • FTO_HUMAN antibody
    • GDFD antibody
    • KIAA1752 antibody
    • mKIAA1752 antibody
    • Protein fatso antibody
    see all

Images

  • Western blot - Anti-FTO antibody (ab94482)
    Western blot - Anti-FTO antibody (ab94482)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: FTO knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HEK293 whole cell lysate (20 µg)
    Lane 4: MOLT4 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab94482 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab94482 was shown to specifically recognize FTO in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when FTO knockout samples were examined. Wild-type and FTO knockout samples were subjected to SDS-PAGE.  Ab94482 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-FTO antibody (ab94482)
    Western blot - Anti-FTO antibody (ab94482)
    All lanes : Anti-FTO antibody (ab94482) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Mouse Hippocampus Tissue Lysate
    Lane 3 : Brain (Mouse) Tissue Lysate
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 58 kDa
    Observed band size: 58 kDa
    Additional bands at: 20 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes

Protocols

  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (4)

Publishing research using ab94482? Please let us know so that we can cite the reference in this datasheet.

ab94482 has been referenced in 4 publications.

  • Zhang X  et al. Extracellular vesicle-encapsulated miR-22-3p from bone marrow mesenchymal stem cell promotes osteogenic differentiation via FTO inhibition. Stem Cell Res Ther 11:227 (2020). PubMed: 32522250
  • Jones PH  et al. Over-expression of miR-34c leads to early-life visceral fat accumulation and insulin resistance. Sci Rep 9:13844 (2019). PubMed: 31554925
  • Li H  et al. FTO is involved in Alzheimer's disease by targeting TSC1-mTOR-Tau signaling. Biochem Biophys Res Commun 498:234-239 (2018). WB . PubMed: 29501742
  • Shen GS  et al. The GDF11-FTO-PPAR? axis controls the shift of osteoporotic MSC fate to adipocyte and inhibits bone formation during osteoporosis. Biochim Biophys Acta Mol Basis Dis 1864:3644-3654 (2018). PubMed: 30279140

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Question

8 nd ICC-IF, mouse samples
No staining
Can we use ab94482 and ab106386 in IHC-Fr, IHC-P and ICC-IF?

Read More

Abcam community

Verified customer

Asked on May 29 2012

Answer

rll yesterday afternoon. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that ab92821 is tested and covered by our 6 month guarantee forICC-IF and mouse. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I can suggest to provide just the information regarding ICC-IF to begin with, as this is covered by the guarantee. As discussed on the telephone, I am sorry to confirm that this antibody is not tested and guaranteed for IHC-P or IHC-Fr. All tested and guaranteed applications and species are listed on the online datasheets.

I would appreciate if you could also provide an image which would help us to assess the results.

With regards to ab94482 and ab106386:

ab94482 is not tested in IHC-Fr, IHC-P and ICC-IF. It has been tested on mouse.

ab106386 is tested in IHC-P only and I am sorry this is not tested in ICC-IF, IHC-Fr or on mouse samples. I have checked the alignment of the humanimmunogen with the mouse protein and this is86%. This is high,so theoreticallythe antibody islikely to detect in mouse. But please note this is not guaranteed because not tested.

I can offer a discount off a future purchase if you buy ab94482and want to testit inin IHC-Fr, IHC-P or ICC-IF. I can also offer a discount of a future purchase in you purchase ab106383 to test in mouse and in IHC-Fror ICC-IF.

I would suggest to wait until the resolution of this case before deciding how you would like to proceed with the discounts. However, I am pleased to provide information on how the discount works.

If you purchase ab94482 or ab106383 to test, then try the antibody and submit feedback to us in the form of an Abreview. It doesn't matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody of the next order after submission of the Abreview. If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed. I will then send a discount code. This code must be issued before purchasing the antibodies so please wait for my reply before ordering. One code will be issued for each antibody.

2. Purchase the antibodieseither by phone, fax, or online (www.abcam.com).

3. Test the antibody in the species and applications discussed above.

ab94482 testin IHC-Fr, IHC-P or ICC-IF.
ab106383 test in mouse and in IHC-Fror ICC-IF.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody] ordered and the discount code is valid for 4 months after issue.

Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the antibody turns out to be unsuitable for the species and application you are testing, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


Order Details
Antibody code:

Lot number:

Purchase order number
or preferably Abcam order number:


General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, non-specific satining etc.)

Sample (Species/Tissue/Cell Type/Cell Line etc.)


Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


Permeabilization step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)


Optimization attempts (problem solving)
How many times have you tried the IHC?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No


What steps have you altered?


Additional Notes


We would appreciate if you are also able to provide and image which would help us to assess the results

Read More

Abcam Scientific Support

Answered on May 29 2012

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