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Rabbit Recombinant Monoclonal Otoferlin antibody. Suitable for IHC-P, WB, IHC-Fr and reacts with Transfected cell line - Mouse, Rat, Mouse samples.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (AB309197), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (AB309197), expandable thumbnail
  • Western blot - Anti-Otoferlin antibody [EPR28223-1] (AB309197), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (AB309197), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (AB309197), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBIHC-FrIP
Human
Not recommended
Not recommended
Not recommended
Not recommended
Mouse
Expected
Tested
Tested
Not recommended
Rat
Tested
Tested
Tested
Not recommended
Transfected cell line - Mouse
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Transfected cell line - Mouse
Dilution info
1/500
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Rat
Dilution info
1/500
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Human, Transfected cell line - Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/100
Notes

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species
Rat
Dilution info
1/100
Notes

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Not recommended
Not recommended

Species
Transfected cell line - Mouse, Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Transfected cell line - Mouse, Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

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.

Alternative names

Fer1l2, Otof, Otoferlin, Fer-1-like protein 2, Protein pachanga

Recommended products

Rabbit Recombinant Monoclonal Otoferlin antibody. Suitable for IHC-P, WB, IHC-Fr and reacts with Transfected cell line - Mouse, Rat, Mouse samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR28223-1
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The otoferlin protein also known as OTOF plays an important mechanical role in the auditory system by serving as an important synaptic vesicle component for neurotransmitter release in inner hair cells of the cochlea. It has a molecular mass of approximately 230 kDa. Otoferlin is expressed primarily in the cochlea particularly in the sensory hair cells essential for hearing. Its functionality hinges on its capacity to serve as a calcium-sensor facilitating the rapid exocytosis of neurotransmitter-filled vesicles at the synaptic ribbon.

Biological function summary

Otoferlin contributes to sound perception by coordinating the timely release of synaptic vesicles that transmit auditory signals to the brain. It is part of a complex network within the auditory pathway interacting with other proteins important for synaptic function including synaptotagmin. Significant for its role in calcium-binding otoferlin participates in the fine-tuning of synaptic transmission ensuring efficient signaling required for precise auditory processing.

Pathways

Otoferlin is integral to the auditory signaling pathways especially the cochlear nerve pathway responsible for conveying sound information to the brain. It actively participates in vesicular trafficking and synaptic vesicle cycle ensuring high fidelity auditory signal transduction. Otoferlin works closely with proteins such as synaptophysin and syntaxin which are involved in the vesicle fusion process highlighting its role in facilitating efficient neurotransmitter release.

Associated diseases and disorders

Otoferlin has a significant association with auditory processing conditions. Mutations in the otoferlin gene lead to severe-to-profound hereditary hearing loss specifically a form of non-syndromic autosomal recessive hearing impairment (DFNB9). Through this condition otoferlin has connections to proteins involved in synaptic vesicle recycling malfunctions contributing to auditory neuropathy. Additionally otoferlin-related dysfunction implicates it in conditions affecting synaptic transmission stability which are vital considerations in therapeutic strategies for hearing loss.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling Otoferlin with ab309197 at 1/500 (1.072 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on (A) HEK-293T transfected with a mouse Otoferlin expression vector containing a his tag. No staining on (B) HEK-293T transfected with empty vector containing a his tagThe section was incubated with ab309197 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Otoferlin with ab309197 at 1/500 (1.072 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Negative control: no staining on mouse liver. The section was incubated with ab309197 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Western blot - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Western blot - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    Negative control: liver (PMID: 10192385).
    In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
    Exposure time: 180 seconds

    Lane 1: Mouse brain tissue lysate at 50 µg

    Lane 2: Mouse liver tissue lysate

    Observed band size: 250 kDa

    Exposure time: 180s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Otoferlin with ab309197 at 1/500 (1.072 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Negative control: no staining on rat liver. The section was incubated with ab309197 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Immunohistochemical analysis of paraffin-embedded Rat cochlea tissue labeling Otoferlin with ab309197 at 1/500 (1.072 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on rat cochlea inner hair cells. The section was incubated with ab309197 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Frozen sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Immunohistochemistry (Frozen sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cochlea tissue labeling Otoferlin with ab309197 at 1/100 (5.36 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining in hair cells on mouse cochlea. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309197 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Immunohistochemical analysis of paraffin-embedded Mouse cochlea tissue labeling Otoferlin with ab309197 at 1/500 (1.072 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Positive staining on mouse cochlea inner hair cells (PMID: 17055430). The section was incubated with ab309197 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/Dab (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).
    Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

  • Immunohistochemistry (Frozen sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Immunohistochemistry (Frozen sections) - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cochlea tissue labeling Otoferlin with ab309197 at 1/100 (5.36 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining in hair cells on rat cochlea. The nuclear counterstain was DAPI (Blue). The section was incubated with ab309197 for 60 mins at room temperature. The section was then mounted using Fluoromount®. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Western blot - Anti-Otoferlin antibody [EPR28223-1] (ab309197), expandable thumbnail

    Western blot - Anti-Otoferlin antibody [EPR28223-1] (ab309197)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    Negative control: liver (PMID: 10192385).
    In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] ab129002) loading control staining at 1/10000 dilution.
    Exposure time: 180 seconds

    All lanes: Western blot - Anti-Otoferlin antibody [EPR28223-1] (ab309197) at 1/1000 dilution

    Lane 1: Rat brain tissue lysate at 50 µg

    Lane 2: Rat liver tissue lysate at 50 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 250 kDa

    Exposure time: 180s

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Product protocols

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