Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Tyrosine Hydroxylase (phospho S31) with ab314484 at 1/6000 (0.086 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on rat cerebrum without alkaline phosphatase treatment (A). No signal was detected when tissues were treated with alkaline phosphatase (B). The section was incubated with ab314484 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Immunohistochemical analysis of paraffin-embedded Mouse adrenal gland tissue labeling Tyrosine Hydroxylase (phospho S31) with ab314484 at 1/6000 (0.086 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse adrenal gland without alkaline phosphatase treatment (A). No signal was detected when tissues were treated with alkaline phosphatase (B). The section was incubated with ab314484 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Tyrosine Hydroxylase (phospho S31) with ab314484 at 1/6000 (0.086 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on mouse cerebrum without alkaline phosphatase treatment (A). No signal was detected when tissues were treated with alkaline phosphatase (B). The section was incubated with ab314484 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) treated with phosphatase at 37℃ overnight (Left) / Untreated PC-12 (Right) cells labelling Tyrosine Hydroxylase (phospho S31) with ab314484 at 1/500 dilution (0.1 ug)/Red and Green (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Tyrosine Hydroxylase (phospho S31) was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab314484 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314484 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse brain tissue lysate Lane 2 : ab314484 IP in Mouse brain tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314484 in mouse brain tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (ab314484) at 1/30 dilution

All lanes:

Mouse brain tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

• IP

Supplier Data

Immunoprecipitation - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Tyrosine Hydroxylase (phospho S31) was immunoprecipitated from 0.35 mg Rat brain tissue lysate with ab314484 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314484 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Rat brain tissue lysate Lane 2 : ab314484 IP in Rat brain tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314484 in rat brain tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (ab314484) at 1/30 dilution

All lanes:

Rat brain tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

• WB

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Western blot - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : liver In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. In Western blot, Anti-Tyrosine Hydroxylase antibody staining at 1/1000 dilution

All lanes:

Western blot - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (ab314484) at 1/1000 dilution

Lane 1:

Mouse adrenal gland tissue lysate (untreated membrane) at 20 µg

Lane 2:

Mouse liver tissue lysate (untreated membrane) at 20 µg

Lane 3:

Mouse adrenal gland tissue lysate (phosphatase treated membrane) at 20 µg

Lane 4:

Mouse liver tissue lysate (phosphatase treated membrane) at 20 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 1/100000 dilution

Observed band size: 60 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. In Western blot, Anti-Tyrosine Hydroxylase antibody staining at 1/1000 dilution

All lanes:

Western blot - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (ab314484) at 1/1000 dilution

Lane 1:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

PC-12 whole cell lysate (phosphatase treated membrane) at 20 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 1/100000 dilution

Observed band size: 60 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : liver. The identity of the band around 37 kDa is unknown. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (ab314484) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse striatum tissue lysate at 20 µg

Lane 3:

Mouse adrenal gland tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Lane 6:

Rat striatum tissue lysate at 20 µg

Lane 7:

Rat adrenal gland tissue lysate at 20 µg

Lane 8:

Rat liver tissue lysate at 20 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 1/100000 dilution

Observed band size: 60 kDa

false

Exposure time: 15s

• Dot

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Dot Blot - Anti-Tyrosine Hydroxylase (phospho S31) antibody [EPR26192-116] (AB314484)

Dot blot analysis of Tyrosine Hydroxylase (phospho S31) using ab314484 at 1 : 1000 (0.514 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Exposure time : 180 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST