Anti-MHC class I + HLA B antibody [EP2624] (ab110645)

Overview

  • Product name
    Anti-MHC class I + HLA B antibody [EP2624]
  • Description
    Rabbit monoclonal [EP2624] to MHC class I + HLA B
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human HLA B. The exact sequence is proprietary.

  • Positive control
    • Ramos, HL60, Jurkat and TF1 cell lysates Paraffin embedded Human tonsils
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab110645 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 1/1000 - 1/10000. Predicted molecular weight: 40 kDa.
IP 1/10 - 1/100.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    Images

    • All lanes : Anti-MHC class I + HLA B antibody [EP2624] (ab110645) at 1/1000 dilution

      Lane 1 : Ramos cell lysate
      Lane 2 : HL 60 cell lysate
      Lane 3 : Jurkat cell lysate
      Lane 4 : TF1 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

      Predicted band size: 40 kDa

    References

    ab110645 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-10 of 13 Abreviews or Q&A

    Answer

    Thank you for your reply.

    I am sorry that this antibody did not perform as stated on the datasheet. As requested, I have asked our Finance department to issue a credit note for you.

    Credit note ID: #####

    The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    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 taking the time to provide the requested details.

    The details you have kindly provided will provide us with vital information for our monitoring of 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 from the alternative antibody. Reviewing the details, it is unusual that the alternative has also not worked and I am sorry there very few tips to provide on this occasion to help improve the results.

    I am pleased to provide a credit note in compensation as requested. In order to do so, I would appreciate if you could confirm that this vial of ab109089 was the alternative free of charge replacement for ab110645 provided in order number 1122795. If not, I would appreciate if you are able to provide the order number and date of purchase.


    Thank you for your cooperation. I look forward to hearing from you with the requested details and hope we can resolve this as soon as possible.

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    Answer

    Thank you for your reply and for kindly confirming these details.

    I understand your concerns and it is regrettable the results have not been successful from this replacement vial aswell.

    As there have been difficulties with two different vials of antibody, I would appreciate if it is possible to investigate this further. This will be helpful information for our quality monitoring, and also I hope we may be able to provide some suggestions to help improve the results:

    1. Could you confirm how the samples have been prepared? Have these cells been cultured at all? Or have the cells been isolated and spun onto a slide straight away for staining (how have they been isolated?)

    2. Has a positive control been included, for example human tonsil as shown on the datasheet?

    3. How long was the fixation in PFA?

    4. Have different antigen retrieval been tried?
    As I am not sure how the samples have been prepared, so I am not currently sure if antigen retrieval is necessary. This is only usually required if the sample has been fixed in PFA for 24 hours.

    Sometimes it is possible to optimize antigen retrieval, and optimization can be different for different epitopes. For example, see the following publication for further information:

    Emoto et al., J Histochem Cytochem. 2005 53 (11) : 1311-21.

    5. Could you confirm the wash steps? We recommend 4 X for 5 minutes at each wash step in PBS containing 0.2% Tween 20. This may help to wash away any non specifically bound antibody.

    6. Is the secondary antibody working well with other primary antibodies?

    7. I can recommend to store the antibody at -20oC as soon as possible, as described on datasheet. This will ensure the antibody remains stable and we can continue to provide our guarantee. Was the antibody diluted before storage, you mention reconstitution?

    I hope this information will be helpful. Please do not hesitate to get back to me with the requested information and I hope we can resolve the case as soon as possible.

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    Answer

    Thank you very much for bringing this to our attention.

    As you point out, this antibody should be referred as anti HLA-B antibody, as it is not specific to allele B07.

    There might have been a mistake when transferring the datasheet to our catalogue from http://www.epitomics.com/products/product_info/1670/HLA-B-antibody-2389-1.html.

    I have passed this information to my colleagues to amend this error as soon as possible.

    We really appreciate feedback from our customers to continuously improve our catalogue. Please do not hesitate to contact us for further assistance.

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    Answer

    Thank you for taking the time to contact us. Your message has been forwarded to me as my colleague Claudia is no longer in the office. I have now taken on this case.

    I am sorry to hear you have also had difficulty obtaining satisfactory results from this replacement antibody. Could you confirm this is ab109089HLA B7 antibody [EPR2623(2)]?

    I would like to reassure you that this antibody is tested and covered by our 6 month guarantee forIHC-Pand 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 would appreciate if you are also able to providethe images, as suggested, as this would help us to assess the results.

    Thank you very much for your continuedtime and cooperation. We look forward to receiving the completed questionnaire.

    Order Details

    Antibody code:

    Lot number:

    Purchase order number
    or preferably Abcam order number:


    General Information
    Antibody storage conditions (temperature/reconstitution etc)


    Description of the problem (high background, low signal, non-specific satining etc.)

    Sample (Species/Tissue/Cell Type/Cell Line etc.)


    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)


    Antigen retrieval (Enzymatic method, Heat mediated technique etc.)


    Permeabilization step


    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


    Positive and negative controls used (please specify)


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



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

    Do you obtain the same results every time?
    Yes No


    What steps have you altered?


    Additional Notes


    We would appreciate if you are also able to provide and image which woudl help us to assess the results

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    Answer

    Thank you for your reply and sorry for the delay in my responce.

    Unfortunately, we are not able to supply sample vials of any or our products. This is due to the size of our catalogue and the logistics this would involve. It simply isn't feasible for us.

    Additionally, as the B27 sequence and the B7 sequences are so similar, 93% homology,(SwissProt reference http://www.uniprot.org/uniprot/P03989and http://www.uniprot.org/uniprot/P01889respectively), I would not think that the antibodies we have would be likely to be able to detect the difference.

    I am sorry that I could not be of more help in this instance. If you have any further questions, please do not hesitate to contact us again.

    Read More

    Answer

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

    Having searched through the antibodies we have against the targets you have mentioned (HLA B7 and B27) we do not have any antibodies which are specific for only one subtype. For example, the immunogen used to raise the anti-HLA B7 antibody [EPR2623(2)], ab110645, shares high homology with multiple subtypes including B-7 and B-27. The same case is true for ab109089, although it is not expected to detect B-27. We have not tested the cross reactivity specifically and therefore cannot be sure exactly which subtype the antibodies is likely to cross react with.

    Similarly, the immunogen used to raise the anti-HLA B27 antibody, ab93150, shares 100% homology with B47, B40, B37, B44 and B13.

    In summary, we cannot guarantee that the antibodies would not cross react, in fact we would expect them to.

    I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

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    Answer

    Just to update you: we determined that the ab110645 is indeed specific for HLAB and not only B7. We will inform all the customers and update the datasheet. Thank you for having drawn our attention to this fact and please accept again our apologies for this problem.

    How are you getting on with the ab109089? Do you obtain the expected results with this antibody?

    I look forward to hear back from you.

    Read More

    Answer

    Thank you foryour fast replyconfirming these details and for your cooperation.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As said, we are investigating this further.

    As requested, I have issued a free of charge replacement of the alternative antibody ab109089 with the order number xx.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Answer

    Thank you for contacting us.

    I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    I have asked the laboratory to retest this antibody. Can you - just to be complete - pleaseanswer the below questions for the laboratory? Thank you very much!

    We will be happy to replace or credit/refund this antibody for you. I apologize for the inconvenience. Please let me know if you prefer to receive a credit/refund or an alternative antibody from our catalog as a replacement. As the order has been placed through xx, we will also need to liaise with with Lucerna in regards on how to provide the replacement/credit. Can you please provide your order number as well?

    Thank you for your cooperation. I look forward to receiving your reply.


    Is this your first time using this antibody?

    How is the product currently being stored?


    Was any pretreatment done on the sample? If yes, please briefly describe the pretreatment reagent and method used (e.g. 10mM Citrate Buffer in 20 minutes in Microwave, 98C)

    Dilution used:
    Type of dilution buffer used:
    Length of incubation with primary:
    Temperature of incubation:
    Volume of primary antibody used (µl)?

    What blocking buffer/substrate was used, how long, at what temperature?
    (e.g. 10% normal goat serum, overnight at 4C)


    Type of secondary (i.e. goat-anti-Rb-HRP, Polymer, goat-anti-Rb-FITC) and supplier:
    Length of incubation:
    Temperature of incubation:
    Dilution used:
    Type of dilution buffer used:

    Read More

    1-10 of 13 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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