Tryptophan hydroxylase 2 Antibody - BSA Free

Novus Biologicals | Catalog # NB100-74555

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Primate, Rabbit

Cited:

Human, Mouse, Rat, Primate

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunohistochemistry Free-Floating, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry Free-Floating, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Tryptophan Hydroxylase 2 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide make to an internal portion of the mouse Tryptophan hydroxylase 2 protein (between amino acids 10-75) [UniProt Q8CGV2]

Reactivity Notes

Human reactivity is weak. Mouse reactivity reported in scientific literature (PMID: 16581041). Rat reactivity reported in scientific literature (PMID: 24066056).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Tryptophan hydroxylase 2 Antibody - BSA Free

Immunohistochemistry-Paraffin: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunohistochemistry-Paraffin: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunohistochemistry-Paraffin: Tryptophan hydroxylase 2 Antibody [NB100-74555] - IHC analysis of a formalin fixed paraffin embedded tissue section of mouse brain using Tryptophan hydroxylase 2 antibody at 1:25 dilution. The staining was developed using HRP-DAB detection method and the nuclei were counterstained with hematoxylin. The antibody generated staining of the neurons.
Western Blot: Tryptophan hydroxylase 2 AntibodyBSA Free [NB100-74555]

Western Blot: Tryptophan hydroxylase 2 AntibodyBSA Free [NB100-74555]

Western Blot: Tryptophan hydroxylase 2 Antibody [NB100-74555] - Total protein from mouse brain, stomach and human brain was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-Tryptophan Hydroxylase 2 in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.
Immunocytochemistry/ Immunofluorescence: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunocytochemistry/ Immunofluorescence: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Tryptophan-hydroxylase-2-Antibody-Immunocytochemistry-Immunofluorescence-NB100-74555-img0011.jpg
Immunocytochemistry/ Immunofluorescence: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunocytochemistry/ Immunofluorescence: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunocytochemistry/Immunofluorescence: Tryptophan hydroxylase 2 Antibody [NB100-74555] - Analysis in mouse brain cells. Image courtesy of product review by Assaf Vestin.
Immunohistochemistry Free-Floating: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunohistochemistry Free-Floating: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunohistochemistry Free-Floating: Tryptophan hydroxylase 2 Antibody [NB100-74555] - PFA-perfused mouse midbrain serotoninergic neurons. AlexaFluor488 used as secondary antibody. Neurons pseudocolored in yellow hot LUT. Image from verified customer review.
Immunoprecipitation: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunoprecipitation: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555]

Immunoprecipitation: Tryptophan hydroxylase 2 Antibody [NB100-74555] - Analysis of raphe nuclei isolated from Sprague-Dawley rat brains using TPH2 Polyclonal Antibody. PA1-778 was coupled to magnetic beads and added to raphe nuclei whole cell extracts for 15 minutes at RT. Immunoprecipitated proteins were assessed by Western blotting (WB), using either mouse anti-TPH2 (Ms TPH2, Lane 2) or sheep anti-TPH2 antibodies (Sh TPH2, Lane 3). As a positive control for the Western blot, non-immunoprecipitated brain lysates were probed with the mouse anti-TPH2 antibody (Lane 1). As a specificity control, a blot containing immunoprecipitated proteins was probed with an antibody specific for Tyrosine Hydroxylase (TH, Lane 4). Data courtesy of the Innovators Program.
Tryptophan hydroxylase 2 Antibody - BSA Free

Immunohistochemistry: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555] -

The 5-HT and TPH2 in the raphe nuclei of the brain. The 5-HT- and TPH2-positive double stained cells in the dorsal raphe nuclei (A). Representative photomicrographs were taken at magnifications of 100×. Quantification of 5-HT and TPH2 (B). Data are expressed as the means ± SD (n = 3). *, p < 0.05; **, p < 0.01 compared with the vehicle group. 5-HT; 5-hydroxytryptamine, TPH2; tryptophan hydroxylase, DAPI; 4′,6-diamidine-2-phenylindole dihydrochloride.
Tryptophan hydroxylase 2 Antibody - BSA Free

Western Blot: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555] -

Enhanced serotonergic phenotypes of differentiated HT22 cells. (A) Immunofluorescence staining of SERT (green), 5-HT1aR (orange) and TPH2 (magenta) in hippocampal primary culture. DAPI staining (blue) indicates the nucleus. Scale bar = 50 μm. (B,C) SERT (green), 5-HT1aR (orange) and TPH2 (magenta) immunofluorescence staining (B) and western blotting (C) of HT22 cells grown in normal growth medium and differentiation medium for 3 d. DAPI staining (blue) indicates the nucleus. Scale bar = 25 μm. GAPDH was used as loading control. Data are representative of at least three independent experiments and are quantified using densitometric analysis. Statistical significance: *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36703746), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Tryptophan hydroxylase 2 Antibody - BSA Free

Western Blot: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555] -

Pet-1 enhances the serotonergic phenotype in differentiated HT22 cells. (A) Immunofluorescence staining of Pet-1 (red) in hippocampal primary culture. DAPI staining (blue) indicates the nucleus. Scale bar = 50 μm. (B,C) Pet-1 (red) immunofluorescence staining (B) and western blotting (C) of HT22 cells grown in normal growth medium and differentiation medium for 3 d. DAPI staining (blue) indicates the nucleus. Scale bar = 25 μm. GAPDH is used as loading control. (D,E) Control siRNA and Pet-1 siRNA were transiently transfected in HT22 cells, changed with normal growth medium and differentiation medium for 3 d. Western blotting for analyzing Pet-1 (D) and SERT, 5-HT1aR, and TPH2 (E) expression. GAPDH, vinculin, and beta -actin were used as loading controls. Data are representative of at least three independent experiments and are quantified using densitometric analysis. Statistical significance: *p <.05, and ***p <.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36703746), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Tryptophan hydroxylase 2 Antibody - BSA Free

Western Blot: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555] -

Functional loss of 5-HT1Areceptor on the corticosterone and thermoregulatory responses. After fluoxetine injection for 4 weeks, the protein expression of TPH2 and c-Fos following 15 min of 8-OH-DPAT challenge was determined by western blot analysis in RN (A and B). Plasma corticosterone levels (C) or rectal temperature measurement (D) following 0, 15, 30, or 45 min of challenge with 8-OH-DPAT were analyzed. In addition, the plasma corticosterone levels were determined both pre- and post-exercise (E). The data are expressed as the mean +/- SD (n = 4 or 8/group). *p < 0.05 and **p < 0.01 compared to the saline group, #p < 0.05 and ##p < 0.01 compared to the saline-injected group without 8-OH-DPAT challenge or exercise. Serum cortisol levels of patients with ME/CFS were determined by EIA (F). The data are expressed as the mean +/- SD (n = 10 and 17, respectively). *p < 0.05 compared to the healthy controls Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38191373), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Tryptophan hydroxylase 2 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: Tryptophan hydroxylase 2 Antibody - BSA Free [NB100-74555] -

The 5-HT and TPH2 in the raphe nuclei of the brain. The 5-HT- and TPH2-positive double stained cells in the dorsal raphe nuclei (A). Representative photomicrographs were taken at magnifications of 100×. Quantification of 5-HT and TPH2 (B). Data are expressed as the means +/- SD (n = 3). *, p < 0.05; **, p < 0.01 compared with the vehicle group. 5-HT; 5-hydroxytryptamine, TPH2; tryptophan hydroxylase, DAPI; 4′,6-diamidine-2-phenylindole dihydrochloride. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31906307), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Tryptophan hydroxylase 2 Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

reported in scientific literature (PMID 30581079)

Immunocytochemistry/ Immunofluorescence

1:500

Immunohistochemistry

1:10-1:500

Immunohistochemistry Free-Floating

reported in scientific literature (PMID 26428905)

Immunohistochemistry-Frozen

reported in scientific literature (PMID 30350781)

Immunohistochemistry-Paraffin

1:100

Immunoprecipitation

1:1000

Western Blot

1:1000
Application Notes
IF usage was reported in scientific literature (see Soiza-Reillly M et al, 2011).

Reviewed Applications

Read 3 reviews rated 4.7 using NB100-74555 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Tryptophan Hydroxylase 2

Tryptophan hydroxylase (TPH) catalyzes the 5-hydroxylation of tryptophan, which is the first step in the biosynthesis of indoleamines (serotonin and melatonin) (Martinez et al., 2001). In mammals, serotonin biosynthesis occurs predominantly in neurons which originate in the Raphe nuclei of the brain, and melatonin synthesis takes place within the pineal gland. Although TPH catalyzes the same reaction within the Raphe nuclei and the pineal gland, TPH activity is rate-limiting for serotonin but not melatonin biosynthesis. Serotonin functions mainly as a neurotransmitter, whereas melatonin is the principal hormone secreted by the pineal gland. The activity of TPH is enhanced by phosphorylation by cAMP-dependent protein kinase (PKA) and Ca2+/calmodulin kinase II (CaM K II) (Jiang et al., 2000; Johansen et al., 1996). CaM K II phosphorylates Ser260 which lies within the regulatory domain of TPH (Jiang et al., 2000).

Alternate Names

NTPH, TPH2

Entrez Gene IDs

121278 (Human); 216343 (Mouse); 317675 (Rat)

Gene Symbol

TPH2

UniProt

Q8IWU9 (Human)

Additional Tryptophan Hydroxylase 2 Products

Product Documents for Tryptophan hydroxylase 2 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for Tryptophan hydroxylase 2 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Tryptophan hydroxylase 2 Antibody - BSA Free

Customer Reviews for Tryptophan hydroxylase 2 Antibody - BSA Free (3)

4.7 out of 5
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Showing  1 - 3 of 3 reviews Showing All
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  • Tryptophan hydroxylase 2 Antibody
    Name: Luiz Alexandre Magno
    Application: Immunohistochemistry-Frozen
    Sample Tested: midbrain
    Species: Mouse
    Verified Customer | Posted 07/19/2018
    The image shows midbrain serotoninergic neurons from mice. Neurons are pseudocolored in yellow hot LUT
    This is a fantastic antibody to detect TPH2 in free floating slices from PFA-perfused mice. I use it with Alexa Fluor 488 2nd antibody.
    Tryptophan hydroxylase 2 Antibody - BSA Free NB100-74555
  • Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Mouse
    Species: Mouse
    Verified Customer | Posted 12/27/2011
  • Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Brain
    Species: Mouse
    Verified Customer | Posted 10/25/2010
    Tryptophan hydroxylase 2 Antibody - BSA Free NB100-74555

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Protocols

View specific protocols for Tryptophan hydroxylase 2 Antibody - BSA Free (NB100-74555):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in 5% BSA and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

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