Overview

  • Product name
    Anti-Otoferlin antibody [13A9]
  • Description
    Mouse monoclonal [13A9] to Otoferlin
  • Host species
    Mouse
  • Specificity
    This antibody reacts specifically with human Otoferlin protein (220 kDa).
  • Tested applications
    Suitable for: IHC-Fr, Flow Cyt, IHC-FoFr, WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Tagged fusion protein, corresponding to amino acids 1-395 of Human Otoferlin.

Properties

Applications

Our Abpromise guarantee covers the use of ab53233 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
IHC-Fr Use at an assay dependent concentration. PubMed: 23650376
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

IHC-FoFr 1/50. PubMed: 19417007
WB 1/200 - 1/1000. Predicted molecular weight: 227 kDa.
IP 1/50 - 1/200.
ICC/IF 1/100 - 1/500.

Target

  • Function
    Key calcium ion sensor involved in the Ca(2+)-triggered synaptic vesicle-plasma membrane fusion and in the control of neurotransmitter release at these output synapses. Interacts in a calcium-dependent manner to the presynaptic SNARE proteins at ribbon synapses of cochlear inner hair cells (IHCs) to trigger exocytosis of neurotransmitter. Also essential to synaptic exocytosis in immature outer hair cells (OHCs). May also play a role within the recycling of endosomes.
  • Tissue specificity
    Isoform 1 and isoform 3 are found in adult brain. Isoform 2 is expressed in the fetus and in adult brain, heart, placenta, skeletal muscle and kidney.
  • Involvement in disease
    Defects in OTOF are the cause of deafness autosomal recessive type 9 (DFNB9) [MIM:601071]. DFNB9 is a form of sensorineural hearing loss. Sensorineural deafness results from damage to the neural receptors of the inner ear, the nerve pathways to the brain, or the area of the brain that receives sound information.
    Defects in OTOF are a cause of non-syndromic auditory neuropathy autosomal recessive (NSRAN) [MIM:601071]. NSRAN is a form of sensorineural hearing impairment with absent or severely abnormal auditory brainstem response but normal otoacoustic emissions. Auditory neuropathies result from a lesion in the area including the inner hair cells, connections between the inner hair cells and the cochlear branch of the auditory nerve, the auditory nerve itself and auditory pathways of the brainstem. In some cases NSRAN phenotype can be temperature sensitive.
  • Sequence similarities
    Belongs to the ferlin family.
    Contains 4 C2 domains.
  • Cellular localization
    Cytoplasmic vesicle > secretory vesicle > synaptic vesicle membrane. Basolateral cell membrane. Endoplasmic reticulum membrane. Cell membrane. Detected at basolateral cell membrane with synaptic vesicles surrounding the ribbon and at the presynaptic plasma membrane in the inner hair cells (IHCs). Colocalizes with GPR25 and RAB8B in inner hair cells.
  • Information by UniProt
  • Database links
  • Alternative names
    • AUNB1 antibody
    • Deafness, autosomal recessive 9 antibody
    • DFNB6 antibody
    • DFNB9 antibody
    • Fer 1 like protein 2 antibody
    • Fer-1-like protein 2 antibody
    • FER1L2 antibody
    • NSRD9 antibody
    • Otof antibody
    • OTOF_HUMAN antibody
    • Otoferlin antibody
    see all

Images

  • Overlay histogram showing SHSY-5Y cells stained with ab53233 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab53233, 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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ICC/IF analysis of mouse inner ear tissue labeling Otoferlin with ab181781. At 30 days postnatal, Otoferlin is detected in the hair cells of the crista ampullaris (CA) , utricular macula (UM) and saccularmacula (SM).

References

This product has been referenced in:
  • Al-Moyed H  et al. A dual-AAV approach restores fast exocytosis and partially rescues auditory function in deaf otoferlin knock-out mice. EMBO Mol Med 11:N/A (2019). Read more (PubMed: 30509897) »
  • Wang F  et al. Establishment of a Gentamicin Cochlear Poisoning Model in Guinea Pigs and Cochlear Nerve Endings Recognition of Ultrasound Signals. Med Sci Monit 24:9429-9435 (2018). Read more (PubMed: 30592260) »
See all 23 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Head)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA buffer pH 7.8
Permeabilization
No
Specification
Head
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 24 2018

Application
IHC - Wholemount
Sample
(Paddlefish) Polyodon spathula Tissue (hair cells in otic vesicle)
Specification
hair cells in otic vesicle

Abcam user community

Verified customer

Submitted Jun 22 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Zebrafish Cell (Whole organism)
Specification
Whole organism
Fixative
Paraformaldehyde
Permeabilization
Yes - triton
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Colin Johnson

Verified customer

Submitted Jun 20 2012

Answer

Vielen Dank fürIhre Anfrage.

Ein IHC Webinar wurde bereits im Juni 2011 gehalten. Sie koennen es sich auf unserer Webseite unter https://www.abcam.com/blog/ oder https://www.abcam.com/blog/index.cfm/Webinars anschauen.

Das Buch "Immunohistochemistry" ist verfuegbar als ab94964 fuer ein Preis von €50.00 + Lieferkosten.

Falls Sie weitere Fragen haben oder der Antikoerper ab53233 Ihnen weiterhin Probleme bereitet, sagen Sie mir bitte Bescheid.

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Answer

DISCOUNT CODE: x


Expiration date: dd mm 2012

I am very pleased to hear you would like to accept our offer and test ab53233 in zebrafish. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for zebrafish and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount

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Answer

Thank you for contacting us. Because we carry over 80,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

Asthe product is to be used in an untested species or application, youare eligible for our testing discount program though:

To our knowledge, ab53233 has not been tested in zebrafish yet. However, the immunogen reveals a sequencesimilarity of 73% and therefore this antibody might cross-react.Thus, I can offer a discount off a future purchase if you buy ab53233 now, test it in zebrafish 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.

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 and test ab53233 in zebrafish. I will then send a discount code. This code must be issued before purchasing ab53233 so please wait for my reply before ordering.

2. Purchase ab53233 either by phone, fax, or online (www.abcam.com).

3. Test it in zebrafish.

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 more about our Abreview system, 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.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab53233 turns out to be unsuitable for zebrafish, you will still receive the discount on your next purchase after your Abreview has been submitted.

Also, please find on our website (https://www.abcam.com/zebrafish) and attached a list of all our antibodies which we guarantee to work with zebrafish.

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: https://www.abcam.com/collaborationdiscount

Read More

Answer

Thank you for your inquiry. The immunogen this antibody was raised against is a tagged fusion protein, corresponding to amino acids 1-395 of Human Otoferlin. (Immunogen section on the datasheet). We are not aware that the epitope that is recognized by this antibody was specifically mapped or further narrowed down. I am sorry that I could not provide you with the requested information on this occasion.  

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