Donkey Anti-Rat IgG H&L (DyLight® 550) preadsorbed (ab102261)

Overview

  • Product name

    Donkey Anti-Rat IgG H&L (DyLight® 550) preadsorbed
    See all IgG secondary antibodies
  • Description

    Donkey polyclonal Secondary Antibody to Rat IgG - H&L (DyLight® 550), pre-adsorbed
  • Host species

    Donkey
  • Target species

    Rat
  • Specificity

    By immunoelectrophoresis and ELISA this antibody reacts specifically with rat IgG and with light chains common to other rat immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Less than 1% cross reactivity to bovine, chicken, goat, horse, human, mouse, rabbit and sheep IgG was detected. This antibody may cross react with IgG from other species.
  • Tested applications

    Suitable for: Flow Cyt, ICC, IHC-Pmore details
  • Minimal
    cross-reactivity


    Chicken, Cow, Goat, Horse, Human, Mouse, Rabbit, Sheep more details
  • Conjugation

    DyLight® 550. Ex: 562nm, Em: 576nm

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C.
  • Storage buffer

    Preservative: 0.09% Sodium azide
    Constituents: 0.2% BSA, PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Antiserum was cross adsorbed using bovine, chicken, goat, horse, human, mouse, rabbit and sheep immunosorbents to remove cross reactive antibodies. This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to DyLight® 550.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • General notes

    DyLight® 550 replaces DyLight® 549.
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab102261 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
Flow Cyt 1/50 - 1/200.
ICC 1/50 - 1/500.
IHC-P 1/50 - 1/500.

DyLight® 550 replaces DyLight® 549. The DyLight® 550 excitation is 562nm and the emission is 576nm. DyLight® 550 provides the same orange-to-red fluorescence and photostability as the DyLight® 549 dye. Please refer to the vial in order to check whether the product you received is DyLight® 549 or DyLight® 550. This information however will have no impact on your experiments. For more information please refer to DyLight® 550.

References

This product has been referenced in:

  • Hess DL  et al. Perivascular cell-specific knockout of the stem cell pluripotency gene Oct4 inhibits angiogenesis. Nat Commun 10:967 (2019). Read more (PubMed: 30814500) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for confirming this information and for your help and cooperation with this case.

As requested, I have asked our accounting department to issue you with a credit note. This can then be redeemed against the invoice of a future order.

Credit ID: ####

As usual if you have any further questions regarding this credit note, please contact the accounts department by email at creditcontrol@abcam.com. Please refer to the credit ID number in any correspondence with the accounting department.

I would like to wish the customer good luck with their research. The technical team is always at your service, should you require further expert advice.

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Answer

Thank you for your message and for your efforts and time with this customer.

I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments, and would like to offer a refund or credit note in compensation (providing the product has been purchased in the last 6 months). In order to arrange this, I would appreciate if you could confirm the order number and date of purchase as previously requested.

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

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Answer

Thank you for kindly providing this further information.

I appreciate your cooperation and understand your concerns. It is regrettable the results have not been successful.

Reviewing the details, I would like to provide some suggestions and considerations that I hope will help solve the problem, and also request some further information which will help with our investigations.

1. There are inherent difficulties with antibody detection of recombinant tagged proteins that I can recommend considering. This may be a biological artifact rather than a problem with the antibody. Could you confirm that the protein transfected is full length and includes the full immunogen sequence for the primary antibody?

Also, if the recombinant E6AP-HA protein is tagged, it is possible that the tag is preventing access to the antibody. Could you confirm at which region of the protein the tag has been placed?

2. Could you confirm how the stability of transfection has been assessed?

3. What type of optimization has been tried with the primary antibodies? For example, has a dilution range been tried? As previously discussed, overnight incubation at 4oC can often provide more efficient and specific staining.

4. I can recommend that due to the points described above, it is important to include an endogenous positive control sample for the primary antibodies.

5. I can suggest it would be important to try the secondary antibody with another primary antibody (detecting an endogenous, not recombinant protein) to help assess how well this is working.

Or, has a different secondary antibody been tried on the same samples with the same primary antibodies?

6. We recommend wash steps are 4 times for 5 minutes at each appropriate wash step if this has not already been tried.

I will be pleased to provide a free of charge replacement or credit note if the antibody is not working with the suggestions provided and an endogenous positive control.

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

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