Overview

  • Product name
    Anti-TRP1 antibody [TA99]
    See all TRP1 primary antibodies
  • Description
    Mouse monoclonal [TA99] to TRP1
  • Host species
    Mouse
  • Specificity
    ab3312 is specific for melanomas and does not react with carcinomas and sarcomas.
  • Tested applications
    Suitable for: IHC-Fr, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    SK-MEL-23 Melanoma cell line.

  • Positive control
    • In Western Blot, this antibody gave a positive signal in human skin tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab3312 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 Use at an assay dependent concentration.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 72 kDa (predicted molecular weight: 72 kDa).
ICC/IF Use at an assay dependent concentration. PubMed: 19841138

Target

  • Function
    Oxidation of 5,6-dihydroxyindole-2-carboxylic acid (DHICA) into indole-5,6-quinone-2-carboxylic acid. May regulate or influence the type of melanin synthesized.
  • Tissue specificity
    Pigment cells.
  • Pathway
    Pigment biosynthesis; melanin biosynthesis.
  • Involvement in disease
    Defects in TYRP1 are the cause of albinism oculocutaneous type 3 (OCA3) [MIM:203290]; also known as Rufous oculocutaneous albinism. An autosomal recessive disorder in which the biosynthesis of melanin pigment is reduced in skin, hair, and eyes. Tyrosinase activity is normal and patients have only moderate reduction of pigment. The eyes present red reflex on transillumination of the iris, dilution of color of iris, nystagmus and strabismus. Darker-skinned individuals have bright copper-red coloration of the skin and hair.
  • Sequence similarities
    Belongs to the tyrosinase family.
  • Cellular localization
    Melanosome membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 5 antibody
    • 5,6 dihydroxyindole 2 carboxylic acid oxidase antibody
    • 6-dihydroxyindole-2-carboxylic acid oxidase antibody
    • b-PROTEIN antibody
    • CAS2 antibody
    • Catalase B antibody
    • CATB antibody
    • DHICA oxidase antibody
    • Glycoprotein 75 antibody
    • GP75 antibody
    • Melanoma antigen gp75 antibody
    • OCA3 antibody
    • TRP antibody
    • TRP-1 antibody
    • TRP1 antibody
    • Tyrosinase related protein 1 antibody
    • Tyrosinase-related protein 1 antibody
    • TYRP antibody
    • TYRP1 antibody
    • TYRP1_HUMAN antibody
    • TYRRP antibody
    see all

Images

  • Anti-TRP1 antibody [TA99] (ab3312) at 5 µg/ml + Human skin tissue lysate - total protein (ab30166) at 20 µg

    Secondary
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 72 kDa
    Observed band size: 72 kDa
    Additional bands at: 61 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes
  • ab3312 staining TRP1 in B16 mouse melanoma cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in 2% paraformaldehyde for 30 minutes and permeabilized in 0.1% Triton X-100 prior to blocking in 5% goat serum for 1 hour at 22°C. The primary antibody was diluted 1/200 in 5% goat serum/1% BSA in PBS and incubated with the sample for 1 hour at 22°C. The secondary antibody was Alexa Fluor® 568-conjugated Goat anti-Mouse polyclonal, diluted 1/500.

    See Abreview

  • Paraformaldehyde-fixed, 0.1% tritonx-100 - permeabilized human skin tissue stained for TRP1 (Red) using ab3312 at 1/300 dilution in immunohistochemical analysis. Phalloidin (Green) DAPI (Blue).

    See Abreview

References

This product has been referenced in:
  • Bergam P  et al. ABCB6 Resides in Melanosomes and Regulates Early Steps of Melanogenesis Required for PMEL Amyloid Matrix Formation. J Mol Biol N/A:N/A (2018). Read more (PubMed: 29940187) »
  • Alaga KC  et al. Aberrant Cx43 Expression and Mislocalization in Metastatic Human Melanomas. J Cancer 8:1123-1128 (2017). Read more (PubMed: 28607585) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunocytochemistry
Sample
Human Cultured Cells (Melanoma cells)
Permeabilization
No
Specification
Melanoma cells
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Acetone

Dr. Christian Ostalecki

Verified customer

Submitted May 24 2018

Abreviews
Application
ELISA
Sample
Human Serum (melanoma patient serum)
Specification
melanoma patient serum
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Type
Direct

Frau. Mirjam Fässler

Verified customer

Submitted Sep 12 2017

Application
Immunohistochemistry (Frozen sections)
Blocking step
0.1% triton, 2.5% goat serum, 2% BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 2% · Temperature: 25°C
Sample
Human Tissue sections (Human Skin)
Specification
Human Skin
Permeabilization
Yes - 0.1% tritonx-100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 09 2014

Answer

Thank you for contacting us and your interest in our products.

We have not used the anti-TRP1 antibody [TA99] ab3312 in in vivo assays and would therefore not be able to guarantee the results. I would like to point out that the antibody has 0.1% azide as a preservative which is likely to be detrimental due to its toxicity. Ab80781 is a BSA and azide free preparation of the clone TA99.

We have not confirmed if the preparation of ab3312 or ab80781 does contain any viruses. We do not expect it would but have not confirmed this.

I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (SK-MEL-28)
Specification
SK-MEL-28
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X in PBS

Abcam user community

Verified customer

Submitted Oct 18 2012

Answer

Thank you for confirming these details.

We have added a free of charge unit of ab34165 to your order xx. This free of charge unit has been provided to replace the ab3312.

Please wish the customer all the best of success with his research.

Thank you for your collaboration. Please do not hesitate to contact us again, should you or the customer have any question.

Read More

Question

Dear technical support team:

This customer has purchased ab3312 (Anti-TRP1 antibody [TA99]) and has conducted the wb several times with human sample. The results show wrong band size, therefore, he wants to ask for your help to modify his experiment step, could you please offer any suggestion to help him?

I also attached his image in this letter and his experiment step as follow:

1. Order details:
Abcam product code: ab3312
Antibody storage conditions (temperature/reconstitution etc): +4℃

2. Please describe the problem (high background, wrong band size, more bands, no band etc).
Wrong band size

3. On what material are you testing the antibody in WB?
Species: human
What's cell line or tissue: RPMI 7951 cell line
Cell extract or Nuclear extract: Cell extract
Purified protein or Recombinant protein: total protein

3. The lysate
How much protein was loaded: 25ug
What lysis buffer was used:Tris 50mM,NaCl 150mM,EDTA 10mM,PMSF 1mM,SDS 0.1%,NP40 1%,triton 0.3% in Q-water
What protease inhibitors were used: NO
What loading buffer was used: 5X samplebuffer: 4ml 1.5M Tris-Cl pH6.8,10ml glycerol, 5ml beta-mercaptoethanol, 2g SDS, 1ml 1% bromophenol blue
Phosphatase inhibitors :NO
Did you heat the samples: temperature and time:95℃,10min

4. Electrophoresis/Gel conditions/ Transfer conditions
Reducing or non reducing gel: non reducing gel
Reducing agent: NO
Gel percentage : 10%
Transfer conditions: (Type of membrane, Protein transfer verified): PVDF,

5. Blocking conditions
Buffer: TBST
Blocking agent: milk, BSA, serum, what percentage: milk 5%
Incubation time:1 h
Incubation temperature: room temperature

6. Primary Antibody
Species: mouse
Reacts against:
At what dilution(s) have you tested this antibody:1:1000,1:2000
What dilution buffer was used: TBST
Incubation time: over night
Incubation temperature: 4℃
What washing steps were done: 5min X3

7. Secondary Antibody
Species: goat
Reacts against:
At what dilution(s) have you tested this antibody: 1:1000,1:3000
Incubation time: 1 H
Wash steps: 5min X3
Fluorochrome or enzyme conjugate: HRP
Do you know whether the problems you are experiencing come from the secondary?

8. Detection method
ECl, ECl+, other detection method: ECL

9. Did you apply positive and negative controls along with the samples? Please specify.
Negative control: H1299, JEG, C9 (human ovarian teratoma transformed epidermal carcinoma cell line)
C9 reference: http://www.ncbi.nlm.nih.gov/pubmed/11087069Lu MH, et al. Proc Natl Sci Counc Repub China B. 2000:169-77

10. Optimization attempts
How many times have you tried the Western? 3 times
Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): background clear
Do you obtain the same results every time e.g. are background bands always in the same place? YES
What steps have you altered? Alter Primary Antibody and Secondary Antibody dilution

Could you please help this customer to solve the problem?

Thanks for your kindly help

Best regards

Read More
Answer

Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

The only recommandation I can make for a future test, are to use fresh samples with proteinase inhibitors (to exclude all possibility of protein degradation).

I apologize for the inconvenience and 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
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (B16 mouse melanoma cell line)
Specification
B16 mouse melanoma cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

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

Answer

Unfortunately, the exact epitope recognised by this antibody has not been mapped

Read More

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