Recombinant
RabMAb

Recombinant Anti-Tryptophan Hydroxylase/TPH antibody [EP1311Y] (ab52954)

Overview

  • Product name

    Anti-Tryptophan Hydroxylase/TPH antibody [EP1311Y]
    See all Tryptophan Hydroxylase/TPH primary antibodies
  • Description

    Rabbit monoclonal [EP1311Y] to Tryptophan Hydroxylase/TPH
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Human Tryptophan Hydroxylase/TPH aa 350-450 (C terminal).

  • Positive control

    • THP-1 cell lysate and human urinary bladder carcinoma.
  • General notes

     

     

     This product was previously labelled as Tryptophan Hydroxylase

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52954 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).
IP 1/50.
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt or ICC/IF.
  • Target

    • Tissue specificity

      Isoform 2 seems to be less widely expressed than isoform 1.
    • Pathway

      Aromatic compound metabolism; serotonin biosynthesis; serotonin from L-tryptophan: step 1/2.
    • Sequence similarities

      Belongs to the biopterin-dependent aromatic amino acid hydroxylase family.
      Contains 1 ACT domain.
    • Information by UniProt
    • Database links

    • Alternative names

      • Indoleacetic acid 5 hydroxylase antibody
      • L tryptophan hydroxylase antibody
      • MGC119994 antibody
      • TPH 1 antibody
      • TPH antibody
      • TPH1 antibody
      • TPH1_HUMAN antibody
      • TPRH antibody
      • TRPH antibody
      • Tryptophan 5 hydroxylase 1 antibody
      • Tryptophan 5 monooxygenase 1 antibody
      • Tryptophan 5 monooxygenase antibody
      • Tryptophan 5-hydroxylase 1 antibody
      • Tryptophan 5-monooxygenase 1 antibody
      • Tryptophan hydroxylase 1 antibody
      see all

    Images

    • Anti-Tryptophan Hydroxylase/TPH antibody [EP1311Y] (ab52954) at 1/500 dilution + THP-1 cell lysate at 10 µg

      Secondary
      goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 51 kDa
      Observed band size: 51 kDa

    • ab52954 at 1/50 dilution staining Tryptophan Hydroxylase/TPH in human urinary bladder carcinoma by Immunohistochemistry, Paraffin embedded tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Overlay histogram showing HeLa cells stained with ab52954 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52954, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    References

    This product has been referenced in:

    • Chen DQ  et al. Identification of serum metabolites associating with chronic kidney disease progression and anti-fibrotic effect of 5-methoxytryptophan. Nat Commun 10:1476 (2019). Read more (PubMed: 30931940) »
    • Sun W  et al. Fecal Microbiota Transplantation Can Alleviate Gastrointestinal Transit in Rats with High-Fat Diet-Induced Obesity via Regulation of Serotonin Biosynthesis. Biomed Res Int 2018:8308671 (2018). Read more (PubMed: 30370307) »
    See all 16 Publications for this product

    Customer reviews and Q&As

    1-7 of 7 Abreviews or Q&A

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    Formaldehyde

    Mr. Carl Hobbs

    Verified customer

    Submitted Jan 20 2016

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Specification
    Brain
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    Formaldehyde

    Mr. Carl Hobbs

    Verified customer

    Submitted Jan 08 2016

    Question

    1) Abcam product code ab 52954



    2) Abcam order reference number or product batch number


    3) Description of the problem weak staining



    4) Sample preparation:

    Species mouse

    Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: PFA/formalin fixed paraffin

    Sample preparation

    Positive control rat lung

    Negative control igG



    5) Fixation step

    Yes

    If yes: Fixative agent and concentration 10% neutral buffered formalin

    Fixation time 24 hours

    Fixation temperature room temp



    6) Antigen retrieval method microwave 10mm citric acid buffer



    7) Permeabilization method:

    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?no

    Permeabilizing agent and concentration:





    8) Blocking agent (eg BSA, serum…): bsa and normal horse serum

    Concentration 10% bsa. 2.5% normal horse serum

    Blocking time 1hour bsa . 30mins serum

    Blocking temperature room temp



    9) Endogenous peroxidases blocked?yes

    Endogenous biotins blocked?no



    10) Primary antibody (If more than one was used, describe in “additional notes”) :

    Concentration or dilution 1:50 dilution

    Diluent buffer 1% bsa in 50mm pbs

    Incubation time overnight



    11) Secondary antibody: ImmPress anti rabbit (vector labs)

    Species:ready to use peroxidase linked secondary antibody

    Reacts against:

    Concentration or dilution prediluted

    Diluent buffer

    Incubation time

    Fluorochrome or enzyme conjugate



    12) Washing after primary and secondary antibodies:

    Buffer tbs

    Number of washes 4 x 15minutes



    13) Detection method ImmPress VIP (vector labs)



    14) How many times have you run this staining? Four times

    Do you obtain the same results every time?yes

    What steps have you altered to try and optimize the use of this antibody?

    Exact same condition used with previous ab52954 with different batch number (please refer to my previous email)

    Read More
    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will provide us with vital information for our monitoring of product quality. I would like to reassure you that this antibody is tested and covered by our guarantee for IHC.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there very few tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial.

    I can recommend you may like to consider trying the following options:

    1.) There is a difference between both vial concerning the concentration:

    new lot: 1.5970 mg/ml
    old lot : 2.8950 mg/ml

    I therefore can suggest to increase the amount of antibody that is used. Our lab checked both lots in parallel and found no difference in staining when using both lot at a dilution of 1: 2000 in WB. The IHC was not checked again for the new lot.

    2.) Could you confirm that all other reagents work as before? Is the secondary working?

    Alternatively, or if these tips do not work, I am pleased to offer you a free of charge replacement or credit note in compensation.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question
    Answer

    DISCOUNT CODE: ***
    Expiration date: March 1, 2013
    Value: $337

    I am very pleased to hear you would like to accept our offer and test ab52954 with baboon samples. This code will give you $337 off your next order before the expiration date. To redeem this offer, please submit an Abreview for ab52954 with baboon samples and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

    Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

    The terms and conditions applicable to this offer can be found here: www.abcam.com/abtrial.

    Read More
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain, Raphe Nucleus)
    Permeabilization
    Yes - Triton X-100 0.1%
    Specification
    Brain, Raphe Nucleus
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Apr 26 2012

    Answer

    Thank you for contacting us. The IHC-FrFl Abreview was submitted to us by one of our customer few months back. We had tried to troubleshoot the protocol however we were not able to resolve the issue. Because we do not have the IHC-Fr tested data of our own lab so we were unable to help customer with the problems they were experiencing. Generally the rabbit monoclonal are quite specific however in this case we were unable to find the cause of high background. We are also not sure whether a different protocol is needed or the antibody actually is unsuitable for IHC-FR. So I am sorry we will be unable to provide further information about its specificity in IHC-FR. We however also cannot judge the unsuitability of antibody from just one Abreview. As IHC-Fr is still untested application so I can provide an Abreview testing discount if you are interested in testing this ab again in IHC-Fr. The discount code will give your next antibody free of charge after submitting an IHC-Fr Abreview. We however also recommend using human bladder tissue sections as positive control. On the other hand we also have other antibody ab32821 which we have tested in IHC-Fr and is fully guaranteed to work in IHC-Fr with human or rat tissue sections. Other species are only predicted to react. Let me know if you would like to receive a discount code. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More
    Application
    Immunohistochemistry free floating
    Sample
    Rat Tissue sections (Hindbrain)
    Specification
    Hindbrain

    Abcam user community

    Verified customer

    Submitted Apr 25 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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