Anti-Otoferlin antibody [EPR28223-1]

• IHC-P

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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 (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 (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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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 (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 (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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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 (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 (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-Fr

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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 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 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).

• IHC-P

Supplier Data

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 (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 (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-Fr

Supplier Data

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 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 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).

• IHC-P

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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 (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 (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

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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 (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

false

Exposure time: 180s

• WB

Supplier Data

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 (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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 250 kDa

false

Exposure time: 180s