Anti-TPH2 antibody [EPR19191]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR19191] (AB184505)

Immunohistochemical analysis of paraffin-embedded Mouse brainstem tissue labeling TPH2 with ab184505 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on dorsal raphe cells of mouse brainstem [PMID : 22912815]. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-TPH2 antibody [EPR19191] (AB184505)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse brainstem tissue labeling TPH2 with ab184505 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on dorsal raphe cells of mouse brainstem [PMID : 22912815]. The nuclear counterstain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab150077 secondary antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR19191] (AB184505)

Immunohistochemical analysis of paraffin-embedded Rat brainstem tissue labeling TPH2 with ab184505 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on dorsal raphe cells of rat brainstem [PMID : 22912815]. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-TPH2 antibody [EPR19191] (AB184505)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum tissue labeling TPH2 with ab184505 at 1/500 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2 ug/ml) dilution (Green). Positive staining on mouse cerebrum. is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 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-TPH2 antibody [EPR19191] (AB184505)

Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling TPH2 with ab184505 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. No staining on rat stomach. Counter stained with Hematoxylin.

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TPH2 antibody [EPR19191] (AB184505)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling TPH2 with ab184505 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. No staining on mouse spleen. Counter stained with Hematoxylin.

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

• IP

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Immunoprecipitation - Anti-TPH2 antibody [EPR19191] (AB184505)

TPH2 was immunoprecipitated from 0.35 mg of Mouse brain lysate with ab184505 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184505 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Mouse brain lysate, 10μg (Input).

Lane 2 : ab184505 IP in Mouse brain lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab184505 in Mouse brain lysate.

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-TPH2 antibody [EPR19191] (ab184505)

Predicted band size: 56 kDa

false

• WB

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Western blot - Anti-TPH2 antibody [EPR19191] (AB184505)

Blocking/Dilution buffer : 5% NFDM/TBST.

This target is specifically expressed in DR (dorsal raphe nucleus). WB was tested with midbrain or brainstem because both include the DR zone. The results were consistent with the literature (PMID : 21272616; PMID : 19181488).

All lanes:

Western blot - Anti-TPH2 antibody [EPR19191] (ab184505) at 1/1000 dilution

Lane 1:

Mouse midbrain lysate at 20 µg

Lane 2:

Mouse brainstem lysate at 20 µg

Lane 3:

Mouse kidney lysate at 20 µg

Lane 4:

Mouse spleen lysate at 20 µg

Lane 5:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg

Lane 6:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell 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

Predicted band size: 56 kDa

Observed band size: 56 kDa

false

Exposure time: 3min