Overview

  • Product name

  • Description

    Rabbit polyclonal to Tyrosinase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Sheep, Rabbit, Goat, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Pig, Chimpanzee
  • Immunogen

    A region within synthetic peptide: TQYESGSMDK AANFSFRNTL EGFASPLTGI ADASQSSMHN ALHIYMNGTM, corresponding to internal sequence amino acids 325-374 of Human Tyrosinase

  • Positive control

    • Jurkat whole cell lysate (ab7899). IF/ICC: Malme-3M cell line.

Properties

Applications

Our Abpromise guarantee covers the use of ab61294 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1.25 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.

Target

  • Function

    This is a copper-containing oxidase that functions in the formation of pigments such as melanins and other polyphenolic compounds. Catalyzes the rate-limiting conversions of tyrosine to DOPA, DOPA to DOPA-quinone and possibly 5,6-dihydroxyindole to indole-5,6 quinone.
  • Involvement in disease

    Defects in TYR are the cause of albinism oculocutaneous type 1A (OCA1A) [MIM:203100]; also known as tyrosinase negative oculocutaneous albinism. An autosomal recessive disorder in which the biosynthesis of melanin pigment is absent in skin, hair, and eyes. It is characterized by complete lack of tyrosinase activity due to production of an inactive enzyme. Patients present with a life-long absence of melanin pigment after birth, and manifest increased sensitivity to ultraviolet radiation with predisposition to skin cancer. Visual anomalies include decreased acuity, nystagmus, strabismus and photophobia.
    Defects in TYR are the cause of albinism oculocutaneous type 1B (OCA1B) [MIM:606952]; also known as albinism yellow mutant type. An autosomal recessive disorder in which the biosynthesis of melanin pigment is reduced in skin, hair, and eyes. It is characterized by partial lack of tyrosinase activity. Patients have white hair at birth that rapidly turns yellow or blond. They manifest the development of minimal-to-moderate amounts of cutaneous and ocular pigment. Some patients may have with white hair in the warmer areas (scalp and axilla) and progressively darker hair in the cooler areas (extremities). This variant phenotype is due to a loss of tyrosinase activity above 35-37 degrees C.
  • Sequence similarities

    Belongs to the tyrosinase family.
  • Cellular localization

    Melanosome membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • ATN antibody
    • CMM8 antibody
    • LB24 AB antibody
    • LB24-AB antibody
    • Monophenol monooxygenase antibody
    • OCA1 antibody
    • OCA1A antibody
    • OCAIA antibody
    • Oculocutaneous albinism IA antibody
    • SHEP3 antibody
    • SK29 AB antibody
    • SK29-AB antibody
    • Tumor rejection antigen AB antibody
    • TYR antibody
    • TYRO_HUMAN antibody
    • tyrosinase (oculocutaneous albinism IA) antibody
    • Tyrosinase antibody
    see all

Images

  • ICC/IF image of ab61294 stained Malme-3M cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab61294, 5µg/ml) overnight at +4°C. The secondary antibody (green) was a96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Anti-Tyrosinase antibody (ab61294) at 1.25 µg/ml + Jurkat cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 60 kDa
    Observed band size: 60 kDa

References

This product has been referenced in:

  • Yi WJ  et al. Degraded melanocores are incompetent to protect epidermal keratinocytes against UV damage. Cell Cycle 17:844-857 (2018). Read more (PubMed: 29623762) »
  • Cao J  et al. Tuberous sclerosis complex inactivation disrupts melanogenesis via mTORC1 activation. J Clin Invest 127:349-364 (2017). Read more (PubMed: 27918305) »
See all 4 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of alternative replacement ab738 has been added to your order number ###. This has Abcam order reference ####.

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

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Answer

Thank you for your reply.

The credit note may be redeemed for refund or credit for future purchase. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
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Answer

Thank you for contacting us. Your credit note ID is **********. I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Answer

Thank you for contacting us regarding the specificity of ab61294.

I have brought your finding to the attention of the lab. They are very concerned and have targeted this product for new quality control testing with samples known to and not to express Tyrosinase. We do apologize for the inconvenience and would like to offer you a replacement product or refund for your original purchase. In order to process this, could you please provide me with your Abcamorder confirmation number or PO number used at purchase?

I look forward to your reply and to hearing how you would like to proceed. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

Thank you for your reply.

I am sorry to confirm that we don't have any different lots in stock at the moment. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. Regrettably, I can suggest you have received a bad vial on this occasion.

Therefore, we would be pleased to provide a replacement from the same lot, which will still be covered by our guarantee. Alternatively, I fully understand the customers concerns and if they prefer a credit note in this case, I will be pleased to arrange this for them.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear the customer has some concerns regardingthe 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.

I would like to reassure you that ab61294 is tested and covered by our 6 month guarantee for use inWB and human samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would appreciate if you can confirm some further details so I can investigate further at this stage. This will help us with our quality monitoring and also helpto establish if this antibody is binding non specifically.

1. Could you confirm if fresh negative control 293T cell samples have been tried to rule out any possible cross contamination between the wells of the gel or samples? Is the band in the negative control showing in every experiment?

2. Is the secondary antibody working well with other primary antibodies? I can recommend to include a no primary control which will help to assess if the secondary antibody is binding non specifically.

3. Has the cell line been tested for presence of any cross contaminating cell lines? For example, is there any PCR data for tyronisase for the non transfected 293T cellsamples you are using?

4. Have any other negative control cell lines been tested? It would be highlybeneficial to the investigation of this case if it is possible you have any other negative control sample data.

I hope this information is helpful, thank you for your cooperation. I look forward to receivingthe further requested details.

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had some concerns regarding the results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB and human samples. In the event that a product is not functioning in the tested applications and species 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 would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I have done a literature search to find further information regarding the expression level of Tyrosinase in MDCK and 293T cells. Unfortunately, I was not able to find much information. Do you have any confirmation of this, and how these were selected as a negative control?

I would appreciate if you could also provide an image, including molecular weight markers, which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.


Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asses the results.

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