Anti-TPH2 antibody [EPR25100-29]

• Flow Cyt

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Flow Cytometry - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (Human embryonic kidney epithelial cell) transfected with a human Tryptophan 5-hydroxylase 2 expression vector containing a myc-His tag cells labelling TPH2 with ab288067 at 1/500 dilution (0.1ug)( Right) compared with a Rabbit monoclonal IgG (ab172730) (Left) isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T cells labelling TPH2 with ab288067 at 1/2000 (0.315 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Cytoplasmic staining on (A) HEK-293T cells transfected with a TPH2 expression vector containing a his tag. No staining on (B) HEK-293T cells transfected with empty vector containing a his tag.The section was incubated with ab288067 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 LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T+OE-1169 cells labelling TPH2 with ab288067 at 1/500 (1.26 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody ready to use(Green). Confocal image showing cytoplasmic staining in 293T cells transfected with myc-tagged TPH2 expression vector. is observed. Myc-Tag Mouse mAb (Alexa Fluor^® 647) was used to counterstain tubulin at 1/100 (0.38ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) is ready to use.

• IP

Supplier Data

Immunoprecipitation - Anti-TPH2 antibody [EPR25100-29] (AB288067)

TPH2 was immunoprecipitated from 0.35 mg HEK-293T transfected with Tryptophan 5-hydroxylase 2 expression vector containing a myc-His-tag® whole cell lysate 10 ug with ab288067 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288067 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HEK-293T transfected with Tryptophan 5-hydroxylase 2 expression vector containing a myc-His-tag® whole cell lysate 10 ug

Lane 2 : ab288067 IP in HEK-293T transfected with Tryptophan 5-hydroxylase 2 expression vector containing a myc-His-tag® whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288067 in HEK-293T transfected with Tryptophan 5-hydroxylase 2 expression vector containing a myc-His-tag® whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second

All lanes:

Immunoprecipitation - Anti-TPH2 antibody [EPR25100-29] (ab288067)

Predicted band size: 56 kDa

false

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling TPH2 with ab288067 at 1/2000 (0.315 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Positive staining in neural fibers of mouse cerebrum.The section was incubated with ab288067 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 LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse midbrain (fresh) tissue labeling TPH2 with ab288067 at 1/500 (1.26 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on mouse midbrain is observed. 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of paraffin-embedded Rat midbrain tissue labelling TPH2 with ab288067 at 1/2000 (0.315 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Positive staining in rat midbrain.The section was incubated with ab288067 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 LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse midbrain tissue staining THP2 with ab288067 at a 1/2000 (0.315 ug/ml) dilution, P2Y12 with ab300140 at 1/40000 (0.013 ug/ml) dilution and GFAP with ab218309 at 1/1000 ( 1.325 ug/ml) dilution.

Panel A : merged staining of anti-THP2 (green; Opal™520), anti-P2Y12 (grey; Opal™570) and anti-GFAP (magenta; Opal™690) on mouse midbrain.
Panel B : anti-THP2 staining the serotonergic neurons in mouse midbrain.
Panel C : anti-P2Y12 staining microglia in mouse midbrain.
Panel D : anti-GFAP staining astrocytes in mouse midbrain.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab2888067, ab300140 and ab218309 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat midbrain (fresh) tissue labeling TPH2 with ab288067 at 1/500 (1.26 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Positive staining on rat midbrain is observed. 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of paraffin-embedded Mouse midbrain tissue labelling TPH2 with ab288067 at 1/2000 (0.315 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Positive staining in mouse midbrain.The section was incubated with ab288067 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 LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling TPH2 with ab288067 at 1/2000 (0.315 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Positive staining in neural fibers of rat cerebrum.The section was incubated with ab288067 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 LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling TPH2 with ab288067 at 1/2000 (0.315 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . No staining in mouse kidney.The section was incubated with ab288067 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 LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling TPH2 with ab288067 at 1/2000 (0.315 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . No staining in rat kidney.The section was incubated with ab288067 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 LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-TPH2 antibody [EPR25100-29] (AB288067)

TPH2 was immunoprecipitated from 0.35 mg Mouse midbrain tissue lysate 10 ug with ab288067 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288067 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse midbrain tissue lysate 10 ug

Lane 2 : ab288067 IP in Mouse midbrain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288067 in mouse midbrain tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 32 seconds

All lanes:

Immunoprecipitation - Anti-TPH2 antibody [EPR25100-29] (ab288067)

Predicted band size: 56 kDa

false

• WB

Lab

Western blot - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : kidney (PMID : 12511643).

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-TPH2 antibody [EPR25100-29] (ab288067) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 56 kDa

false

• WB

Lab

Western blot - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : kidney, spleen (PMID : 12511643).

Exposure time : Lanes 1-3 : 70 seconds; Lanes 4-6 : 26 seconds

All lanes:

Western blot - Anti-TPH2 antibody [EPR25100-29] (ab288067) at 1/1000 dilution

Lane 1:

Mouse midbrain tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Rat midbrain tissue lysate at 20 µg

Lane 5:

Rat kidney tissue lysate at 20 µg

Lane 6:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 56 kDa

false

• WB

Lab

Western blot - Anti-TPH2 antibody [EPR25100-29] (AB288067)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 10 seconds

All lanes:

Western blot - Anti-TPH2 antibody [EPR25100-29] (ab288067) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containi a myc-His-tag®, whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with Tryptophan 5-hydroxylase 2 expression vector containi a myc-His-tag®, whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 56 kDa

false