Overview

  • Product name

    Anti-FTO antibody [5-2H10]
    See all FTO primary antibodies
  • Description

    Mouse monoclonal [5-2H10] to FTO
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Horse, Cow, Dog, Non human primates
  • Immunogen

    Synthetic peptide from Human FTO.

  • Positive control

    • WB: Rat testes nuclear lysate. ICC/IF: Neuronal progenitor cells. Flow Cyt: SH-SY5Y cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab92821 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 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
ICC/IF 1/100.
Flow Cyt Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. PubMed: 28931651

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.
  • Information by UniProt
  • Database links

  • 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

  • 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 - ab92821 observed at 58 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab92821 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. ab92821 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Overlay histogram showing SH-SY5Y cells stained with ab92821 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92821, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ICC/IF image of ab92821 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab92821, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • Anti-FTO antibody [5-2H10] (ab92821) at 1/1000 dilution + Rat testes lysate

    Predicted band size: 58 kDa
    Observed band size: 58 kDa

References

This product has been referenced in:

  • Tang X  et al. The role of the fat mass and obesity-associated protein in the proliferation of pancreatic cancer cells. Oncol Lett 17:2473-2478 (2019). Read more (PubMed: 30719115) »
  • Song H  et al. METTL3 and ALKBH5 oppositely regulate m6A modification of TFEB mRNA, which dictates the fate of hypoxia/reoxygenation-treated cardiomyocytes. Autophagy 15:1419-1437 (2019). Read more (PubMed: 30870073) »
See all 12 Publications for this product

Customer reviews and Q&As

1-2 of 2 Q&A

Answer

Thank you for you inquiry.

I am happy to confirm that the concentration of ab92821 is 1mg/ml.

I hope this is helpful and wish a good start into the new week.

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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

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