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

Submit a review Q&A (3)References (3)

Key features and details

  • Donkey Anti-Rat IgG H&L (DyLight® 550) preadsorbed
  • Conjugation: DyLight® 550. Ex: 562nm, Em: 576nm
  • Host species: Donkey
  • Isotype: IgG
  • Suitable for: Flow Cyt, ICC, IHC-P

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 750 Biotin DyLight® 488 DyLight® 650 FITC HRP PE Texas Red ® Unconjugated

Overview

  • Product name

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

  • 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

    pH: 6.8
    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

The Abpromise guarantee

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.
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.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (3)

Publishing research using ab102261? Please let us know so that we can cite the reference in this datasheet.

ab102261 has been referenced in 3 publications.

  • Newman AAC  et al. Multiple cell types contribute to the atherosclerotic lesion fibrous cap by PDGFRβ and bioenergetic mechanisms. Nat Metab 3:166-181 (2021). PubMed: 33619382
  • Hess DL  et al. Perivascular cell-specific knockout of the stem cell pluripotency gene Oct4 inhibits angiogenesis. Nat Commun 10:967 (2019). PubMed: 30814500
  • Durgin BG  et al. Smooth muscle cell-specific deletion of Col15a1 unexpectedly leads to impaired development of advanced atherosclerotic lesions. Am J Physiol Heart Circ Physiol 312:H943-H958 (2017). PubMed: 28283548

Customer reviews and Q&As

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Submit a review Submit a question

1-3 of 3 Abreviews or Q&A

Question

Thank you for the reply.

The customer decided to receive a refund. Please issue Zotal a credit note.

These are the ordering details:

####

Thanks in advance for your assistance.

Have a nice day,

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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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Question

Thank you for the detailed reply.

I met the customer and she explained me that she performed the parallel stainingof the same cells transfected with the same plasmid at the same time. The staining was performed with the same 1ary ab and the only difference was the secondary ab: AlexaFluor in comparison to Dylight.

The staining with Alexa succeeded whereas Dylight showed non specific staining.

Please advise.

Thanks in advance fro your reply.

Bset Regards,

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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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Question

Thank you for the reply.

Please read below customer's details:

1. Yes, this is not specific staining.

I used primary Rat-anti-HA Ab (Roche 12075900)

2. i. C33A were stained in Image004_ch00

primary keratinocytes were stained in Image 6-1_ch00

ii. The target is transfected E6AP-HA protein. Dilution of primary Ab is 1:150

iii. Incubation time is 2h in room temperature for the primary antibody

iv. Antibody is week and non specific.

3. I didn't use this secondary antibody with other primary antibodies.

I will include a no primary control and I can try to lower concentration of the secondary antibody, but I don't believe that it will improve the results since the antibody is very week and since I try to detect the transfected protein, I expect to see only several cells stained (like in Images 005 and 014) and not the whole field weekly stained.

Please advise.

Thank you for the assistance.
Best Regards,

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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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