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Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rat, Primate, Rabbit

Applications

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

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for Tryptophan hydroxylase 2 Antibody - BSA Free

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 [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 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 [NB100-74555] - The 5-HT and Tryptophan hydroxylase 2 in the raphe nuclei of the brain. The 5-HT- and Tryptophan hydroxylase 2-positive double stained cells in the dorsal raphe nuclei. Representative photomicrographs were taken at magnifications of 100x. Quantification of 5-HT and Tryptophan hydroxylase 2. Image collected and cropped by CiteAb from the following publication (https://www.mdpi.com/2218-273X/10/1/71), licensed under a CC-BY license.

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).
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

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

Published Applications

Read 53 publications using NB100-74555 in the following applications:

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

Product Documents for Tryptophan hydroxylase 2 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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.

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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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