Overview

  • Product name
  • Description
    Rabbit polyclonal to TRP2/DCT
  • Host species
    Rabbit
  • Specificity

    ab74073 detects endogenous levels of total TRP2/DCT protein.

  • Tested applications
    Suitable for: IHC-Fr, IHC-P, WB, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human TRP2/DCT (internal sequence). synthesized peptide derived from internal of human TRP2.

  • Positive control
    • WB: Extracts from K562 cells ICC/IF: A549 cells
  • General notes

    Previously labelled as TRP2

Properties

Applications

Our Abpromise guarantee covers the use of ab74073 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
IHC-Fr 1/100.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/500 - 1/1000. Predicted molecular weight: 59 kDa.

TRP2 protein is a single-pass type I membrane protein, these types of proteins known to aggregate when boiled therefore, we recommend heating the lysates in sample buffer for 15-20 minutes at 60-70C instead of boiling.   

ELISA 1/40000.
ICC/IF 1/100 - 1/500.

Target

  • Function
    Involved in regulating eumelanin and phaeomelanin levels.
  • Pathway
    Pigment biosynthesis; melanin biosynthesis.
  • Sequence similarities
    Belongs to the tyrosinase family.
  • Cellular localization
    Melanosome membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • L dopachrome tautomerase antibody
    • DCT antibody
    • Dopachrome delta isomerase antibody
    • Dopachrome tautomerase antibody
    • Dopachrome tautomerase dopachrome delta isomerase, tyrosine related protein 2 antibody
    • DT antibody
    • EC 5.3.3.12 antibody
    • L dopachrome Delta isomerase antibody
    • L-dopachrome Delta-isomerase antibody
    • L-dopachrome tautomerase antibody
    • TRP 2 antibody
    • TRP-2 antibody
    • TRP2 antibody
    • Tyrosinase related protein 2 antibody
    • Tyrosinase-related protein 2 antibody
    • TYRP2 antibody
    • TYRP2_HUMAN antibody
    see all

Images

  • ab74043 staining TRP2/DCT in human skin tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde and a heat mediated antigen retrieval step was performed using a TRIS/EDTA buffer pH 9.0. Samples were then permeabilized, followed by incubation with the primary antibody at a 1/1000 dilution for 30 minutes at 20°C. An undiluted HRP-conjugated goat polyclonal was used as secondary antibody.

  • ab74073 staining TRP2/DCT in Mouse melan-ink melanocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 5% serum for 30 hours at room temperature. Samples were incubated with primary antibody (1/100 in PBS -T+ 1% BSA) for 12 hours. An undiluted Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-TRP2/DCT antibody (ab74073) at 1/500 dilution

    Lane 1 : Extracts from K562 cells with no peptide
    Lane 2 : Extracts from K562 cells with peptide at 5 µg

    Lysates/proteins at 5 µg per lane.

    Predicted band size: 59 kDa
    Observed band size: 59 kDa
    Additional bands at: 117 kDa, 39 kDa. We are unsure as to the identity of these extra bands.

  • Immunofluorescence analysis of TRP2/DCT in A549 cells, using ab74073 at a dilution of 1/500.
    Left hand panel: no peptide
    right hand panel: with immunising peptide.

References

This product has been referenced in:
  • Liu Y  et al. Exosomal miRNA derived from keratinocytes regulates pigmentation in melanocytes. J Dermatol Sci 93:159-167 (2019). Read more (PubMed: 30904353) »
  • Chatatikun M  et al. Anti melanogenic effect of Croton roxburghii and Croton sublyratus leaves in a-MSH stimulated B16F10?cells. J Tradit Complement Med 9:66-72 (2019). Read more (PubMed: 30671368) »
See all 24 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Skin)
Permeabilization
No
Specification
Skin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Ahmar Aziz

Verified customer

Submitted Feb 10 2016

Answer

The K562 cells were not transfected. I am not sure how they were chosen for the western blot test, as I have not found any papers that used K562 to study Trp2. Most of the studies are using melanocytes, for instance the paper at this link: http://www.ncbi.nlm.nih.gov/pubmed/?term=8306979.

Another Trp2 antibody, ab92995, was also validated for western blotting using K562. Here is a link to the datasheet:

https://www.abcam.com/trp2-antibody-ab92995.html .

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (melan-ink melanocytes)
Specification
melan-ink melanocytes
Blocking step
Serum as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Mr. Heiko Locher

Verified customer

Submitted Sep 23 2013

Question
Answer

Thank you very much for your interest our AbTrial program.

To our knowledge, ab74073 has not been tested in FACS. However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting us know you are interested in testing this product.

2. Our scientists will email you an inactive personal discount code for the value of the product.

3. Purchase and test the product at the regular price.

4. Submit your results, including your discount code in the additional notes section of your Abreview.

5. Once the Abreview is submitted, the discount code will become active.

6. Apply your discount code on your next order to receive that value off.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

Read More

Answer

Thank you for contacting Abcam regarding ab74073.


Regarding the specificity of this antibody, it will recognize both endogenous TRP2 as well as exogenous protein (cells transfected with full length TRP2). Please see the customer Abreview for a WB of HEK293 cells transfected with TRP2 on the product datasheet on our website.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (skin)
Permeabilization
Yes - 0.1% TritonX-100
Specification
skin
Blocking step
3% BSA, 5% NGS, 0.2%TritonX-100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 09 2012

Question
Answer


I am very pleased to hear you would like to accept our offer and test ab74073 in IHC-Fr]. This code will give you 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview forIHC-Fr and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: 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/collaborationdiscount.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (B16 mouse melanoma cell line)
Permeabilization
Yes - 0.1% Triton X-100
Specification
B16 mouse melanoma cell line
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Dr. James Harris

Verified customer

Submitted Sep 27 2011

Answer

Thank you for your inquiry. Unfortunately, we do not know whether the light chains in these antibodies (ab3312 and ab74073) are lambda (λ) or kappa (κ). This has not been tested. If you require a secondary antibody for these primary antibodies, I can suggest to choose the secondary according to the heavy chain of the primary. Please follow this link to find our search engine for secondary antibodies: https://www.abcam.com/index.html?pageconfig=productmap&cl=918 ab3312 (Anti-TRP1 antibody [TA99]) is a mouse IgG2a and can be detected with anti mouse IgG2a antibodies or general anti mouse IgG antibodies. ab74073 (Anti-TRP2 antibody) is a rabbit IgG and can be detected with an anti rabbit IgG secondary antibody. We offer many secondary antibodies, pre-absorbed, conjugated and un-conjugated. Please let me know if you need help choosing the right secondary antibody for your purposes. In order to help I will need more information about the application, etc you are planning to do. I hope this information was helpful.

Read More

Question

It will take approximately 5 minutes to complete the questionnaire. Please fill-in all fields so that we can better assist you. 1) Abcam product code ab ab74073 2) Abcam order reference number or product batch number GR441991-1 3) Storage conditions: -20℃ 4) Description of the problem (high background, no band, wrong band size...) No staining 5) Sample preparation: Name of the tissue/cell line: skin Sample species: Mice Type of sample (whole cell lysates, fraction, recombinant protein…): tissue lysates Lysis buffer tissue lysis buffer for Western and IP Protease inhibitors: PMSF Phosphatase inhibitors Reducing agent Boiling for ≥5 min? yes/no yes Protein loaded ug/lane or cells/lane 20ul Positive control: β-actin Negative control: No 6) Percentage of gel: 12% Type of membrane Protein transfer verified: Pure Nitrocellulose Blotting Membrane Blocking agent and concentration: Albumin Bovine 5% Blocking time: 1h Blocking temperature: Room temperature 7) Primary antibody (If more than one was used, describe in “additional notes”) : Concentration or dilution: 1:1000 Diluent buffer: TBST Incubation time: overnight Incubation temperature: 4℃ 8) Secondary antibody: Species: Goat Anti-Rabbit lgG-HRP conjugated secongdary antibody Reacts against: Hu Ms Concentration or dilution: 1:10000 Diluent buffer : TBST Incubation time: 1h Incubation temperature: 37℃ Fluorochrome or enzyme conjugate: HRP 9) Washing after primary and secondary antibodies: Buffer: TBST Number of washes: 4-5 times 10)Detection method Exposure 11) How many times have you run this staining? More than 3 times Do you obtain the same results every time? yes What steps have you altered to try and optimize the use of this antibody? Adjustment the antibody Concentration and Blocking agent Concentration Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

Read More
Answer

Thank you once again 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 enable us to investigate this case for you and this is also helpful in our records for monitoring of quality. Reviewing this case, I would like to offer some suggestions to help optimise the results from ab74073. I would also appreciate if you can confirm some further details: 1. Please confirm if there is no staining at all on the blot, or if there is some background at all. Unless the blot is completely blank I would appreciate if you are able to provide an image, including molecular weight markers, which would help me to assess the results. 2. Could you confirm if the sample is already IP purified? I would appreciate if you could provide details of the IP procedure. 3. Please confirm which lysis buffer has been used? I can recommend to try RIPA if you have not already done so. This should provide a suitable protein preparation. 4. Has a positive control whole cell lysate such as K562 cells or A549 cells been included? I can recommend this would be beneficial. 5. I can suggest it is important to assess how much protein is actually loaded. The protein concentration of the sample should be measured, and we recommend to load 20 - 30 ug of whole cell lysate, or 10 - 100 ng of purified protein per lane of a gel. 6. I can suggest to try a higher concentration of antibody 1:500 in the western blot to increase the signal. 7. Could you confirm the results of the loading control? In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

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1-10 of 12 Abreviews or Q&A

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