702017

antibody from Invitrogen Antibodies

Targeting: TPH2 FLJ37295, NTPH

Antibody data

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Validation data

Product number: 702017 - Provider product page
Provider: Invitrogen Antibodies
Product name: TPH2 Recombinant Rabbit Monoclonal Antibody (9H1L6)
Antibody type: Monoclonal
Antigen: Other
Description: This antibody is predicted to react with Monkey, Sheep and Rat.
Antibody clone number: 9H1L6
Concentration: 0.5 mg/mL

TPH2 protein structure - 702017 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-TPH2 Recombinant Rabbit Monoclonal Antibody (9H1L6) (Product # 702017) and a 56 kDa band corresponding to Tryptophan 5-hydroxylase 2 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of IMR-32 (Figure a, Lane 1), U-87 MG (Figure a, Lane 2), SH-SY5Y (Figure a, Lane 3), Mouse Brain (Figure a, Lane 4), Mouse Cerebellum (Figure a, Lane 5), Mouse Ovary (Figure a, Lane 6), Mouse Heart (Figure a, Lane 7), Rat Brain (Figure a, Lane 8), Rat Heart (Figure a, Lane 9) and U-87 MG (Figure b, Lane 1), HeLa (Figure b, Lane 2) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on tissue extracts (30 µg lysate) of Mouse Brain (Lane 1), Rat Brain (Lane 2) and Mouse cerebellum (Lane 3). The blots were probed with Anti-TPH2 Recombinant Rabbit Monoclonal Antibody (Product # 702017, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 56 kDa band corresponding to TPH2 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis was performed on fixed and permeabilized SH-SY5Y cells for detection of endogenous TPH2 using Anti-TPH2 Recombinant Rabbit Monoclonal Antibody (Product # 702017, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of TPH2 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of TPH2. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Tryptophan 5-hydroxylase 2 was performed using 70 confluent log phase U-87 MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with TPH2 Recombinant Rabbit Monoclonal Antibody (9H1L6) (Product # 702017) at 1:200 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2500), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents low expression model, HeLa. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.