Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed (ab96921)

Submit a review Q&A (6)References (18)
Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed (ab96921)
  • Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed (ab96921)

Key features and details

  • Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed
  • Conjugation: DyLight® 594. Ex: 593nm, Em: 618nm
  • Host species: Donkey
  • Isotype: IgG
  • Suitable for: Flow Cyt, ICC/IF, IHC-P

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 750 Alexa Fluor® 790 Alkaline Phosphatase Biotin DyLight® 488 DyLight® 550 DyLight® 650 FITC Gold 10nm Gold 12nm Gold 25nm Gold 6nm HRP Texas Red ® TRITC Unconjugated Unconjugated

Overview

  • Product name

    Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed
    See all IgG secondary antibodies

  • Host species

    Donkey

  • Target species

    Rabbit

  • Specificity

    By immunoelectrophoresis and ELISA this antibody reacts specifically with Rabbit IgG and with light chains common to other Rabbit immunoglobulins. No antibody was detected against non immunoglobulin serum proteins. Reduced cross-reactivity to bovine, chicken, goat, horse, human, mouse, pig and rat was detected.

     

  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, IHC-Pmore details

  • Minimal
    cross-reactivity


    Chicken, Cow, Goat, Horse, Human, Mouse, Pig, Rat more details

  • Conjugation

    DyLight® 594. Ex: 593nm, Em: 618nm

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, human, mouse, pig and rat immunosorbents to remove cross reactive antibodies. This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to DyLight® 594.

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab96921 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/IF
1/50 - 1/500.
IHC-P
1/50 - 1/500.
Notes
Flow Cyt
1/50 - 1/200.
ICC/IF
1/50 - 1/500.
IHC-P
1/50 - 1/500.

Images

  • Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed (ab96921)
    Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed (ab96921)

    E14 mouse brain sample was stained with primary antibody Rabbit anti-Clim1. The secondary antibody used was ab96921 (1:200 dilution, incubate for 1 hour at room temperature). The blue background was from DAPI.

    This image is courtesy of an anonymous Abreview

  • Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed (ab96921)
    Donkey Anti-Rabbit IgG H&L (DyLight® 594) preadsorbed (ab96921)

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 (18)

Publishing research using ab96921? Please let us know so that we can cite the reference in this datasheet.

ab96921 has been referenced in 18 publications.

  • Liu J  et al. Increased alveolar epithelial TRAF6 via autophagy-dependent TRIM37 degradation mediates particulate matter-induced lung metastasis. Autophagy 18:971-989 (2022). PubMed: 34524943
  • Xiu H  et al. Neddylation Alleviates Methicillin-Resistant Staphylococcus aureus Infection by Inducing Macrophage Reactive Oxygen Species Production. J Immunol 207:296-307 (2021). PubMed: 34183370
  • Rajendran A  et al. Tissue-specific deletion of mouse basolateral uniporter LAT4 (Slc43a2) reveals its crucial role in small intestine and kidney amino acid transport. J Physiol 598:5109-5132 (2020). PubMed: 32841365
  • Dolgodilina E  et al. Choroid plexus LAT2 and SNAT3 as partners in CSF amino acid homeostasis maintenance. Fluids Barriers CNS 17:17 (2020). PubMed: 32046769
  • Oparija-Rogenmozere L  et al. Phosphorylation of mouse intestinal basolateral amino acid uniporter LAT4 is controlled by food-entrained diurnal rhythm and dietary proteins. PLoS One 15:e0233863 (2020). PubMed: 32470053
View all Publications for this product

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Question

Customer inquired about detecting presence of 7 different proteins in brain tissue via IHC-F. Would like 3-4 different host species for primary, secondary antibodies that do not overlap in spectra. Customer is also not sure if Caspase 3, Nogo, and neurofilament are necessary for detecting apoptosis / neuronal response.

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Answer

Thank you for contacting Abcam.

I managed to find antibodies against rat GFAP, FOX3, Iba1, and CD68 all raised in different hosts (chicken, rabbit, goat, and mouse, respectively). They each are highly rated by other researchers and have numerous specific published references. The list for these 4 antibodies is given below:
1. GFAP ab4674 (https://www.abcam.com/gfap-antibody-astrocyte-marker-ab4674.html)
2. FOX3 ab104225 (https://www.abcam.com/fox3neun-antibody-ab104225.html)
3. Iba1 ab5076 (https://www.abcam.com/iba1-antibody-ab5076.html)
4. CD68 ab31630 (https://www.abcam.com/cd68-antibody-ed1-ab31630.html)

I also compiled anti-chicken, rabbit, goat, and mouse secondaries with separate emission spectra. We don't have secondaries for these species that weren't raised in any of the 4 species conjugated to a fluorescent protein with a similar spectrum to DAPI. Thus, I think your best bet is to obtain a far-red filter (Cy5), that way you could still stain with DAPI as well. The donkey secondaries that I found with separate emission spectra are listed below:
1. anti-chicken: FITC (https://www.abcam.com/chicken-igy-secondary-antibody-ab63507.html)
2. anti-rabbit: DyLight 594 (Similar spectrum to Texas Red) (https://www.abcam.com/donkey-polyclonal-secondary-antibody-to-rabbit-igg-h-l-dylight-594-pre-adsorbed-ab96921.html)
3. anti-goat: DyLight 650 (Similar spectrum to Cy5) (https://www.abcam.com/donkey-polyclonal-secondary-antibody-to-goat-igg-h-l-dylight-650-pre-adsorbed-ab96938.html)
4. anti-mouse: DyLight 550 (similar spectrum to TRITC) (https://www.abcam.com/donkey-polyclonal-secondary-antibody-to-mouse-igg-h-l-dylight-550-ab96876.html)

The other antibodies you chose (Nogo, heavy neurofilament, caspase 3) seem to be good choices as well as they are tested in formaldehyde fixed, frozen sections via IHC in rat. Below are the antibodies we discussed against those three proteins:
1. anti-Nogo (https://www.abcam.com/nogo-antibody-ab32298.html)
2. anti-heavy neurofilament (https://www.abcam.com/200-kd-neurofilament-heavy-antibody-ab134459.html)
3. anti-caspase 3 (https://www.abcam.com/active-caspase-3-antibody-ab13847.html)

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

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Question

Here isPO 310-61664B

Best regards,

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Answer

Thank you for your email.

The PO-310-61664B was ordered in October 2011 and the complaint was raised in July 2012; this means the product is out of Abpromise guarantee, which is 6 months from date of purchase.

We have sold many units of these products without receiving any quality complaint so I believe that the problem might be with storage. Were these stored in dark; if not, then conjugates might have been damaged due to photo bleaching. Could you let me know how the products were stored?

Thank you very much for your cooperation. I will look forward to hearing from you soon.

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Question

Hello
Our customer said that primary antibodies are displayed at images (attached again in this email).
In addition, her primary antibodies are raised on goat, so she can not
use Alexa.
Antibodies were stored as recommended (4oC).
Order number - PI-004-61517-br
Looking foward for a answer,
Best regards,

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Answer

Could you please check the purchase order number; the number I-004-61517-br or 004-61517 provided does not match in our system? Could you also provide the purchase date of the product?
I will look forward to hearing from you soon.

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Question

Attached below are the questionnaires of our customer.
She is having problems with their secundary antibodies.
Best regards,

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Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

Could you provide the species of all primary antibodies used?
Did you same experimental procedure with Alexa Fluor antibodies?
Have you checked the Green and Redfilters
Could you provide the order number? Howe the antibodies were stored?

I will be looking forward to hearing from you soon.

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Question

thank You for this quick and precise answer. You are suggesting to use DyLight 594, but I am not entirely sure that it would be well excited by the 543nm line; moreover, this excitation wavelength is not mentioned in the AbCam fluorochrome guide. From Your suggestions, I can see that also TRITC and Texas Red have a good emission separation and are both excited at 543nm. Therefore, I would prefer to use products ab7136 (TRITC-conj. anti-guinea pig) and ab7081 (Texas Red-conj. anti-rabbit). Does this make sense or am I missing something? Thank You again for Your precious help. Merry Christmas,

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Answer

Thank you for your reply! Indeed, firstly I wanted to suggest also to use Texas Red with TRITC. However when I looked closer at the emission spectra, it seemed to me that there is quite a "bleeding" of the TRITC into the Texas Red. However this depends also on the staining intensities and the filters available on the microscope. http://www.mcb.arizona.edu/ipc/fret/index.html The DyLight 594 however is still excited (to about 40%) with the laser at 542nm. I thought therefore this might be the safer option. https://www.abcam.com/ps/CMS/Images/DL_594_orange[1]378px302px.jpg It is however up to you to decide. Either option you will chose, we will continue to support you. We will also reward you with points for discounts/amazon vouchers if you can provide your feedback with an image. I hope this information is helpful. Please do not hesitate to contact us again should you have further questions. Merry Christmas and happy new Year to you as well!

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Question

Dear AbCam scientific support, I wish to perform a double immunofluorescence staining (i.e. using two different primary antibodies) on brain sections coming from GFP-expressing mice. However, my confocal microscope has only two excitation wavelengths (488nm and 543nm) but the spectral bandwidth of detection can be regulated. Thus, I am wondering whether I could use two different secondary antibodies according to the following possible combinations: 1) 488nm excites both GFP and secondary antibody number 1; secondary antibody number 2 is excited at 543nm. 2) 488nm excites GFP only; both secondary antibodies are excited at 543nm. This approach could be feasible if I had two secondary antibodies whose emission peaks could be separated by regulating the detection bandwidth. For this reason, I am writing to ask whether such products are available in Your catalogue. Since I am working on mouse tissue, my primary antibodies would be raised in rabbit and guinea pig, respectively. Thanks for Your valuable help. Kind regards,

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Answer

Thank you very much for your inquiry. Indeed we do have a number of antibodies which can be suitable for your application. Please find here below a selection. I have only proposed pre-absorbed antibodies, as they will help to reduce background in immunohistochemistry, especially when doing double stainings. We do also have other formulations of many of these antibodies. For example, we do have antibodies also produced in donkey, against the Fc region or as an F(ab')2. Please do let me know if you have any preference in these regards and I'll be happy to include them in the selection. I have excluded in this list fluorochromes who could interfere with the GFP staining. If this would not matter however, please let me know. Indeed, the request formulated is rather specific. The best option I can propose is, to use the Cy3.5 conjugated anti guinea pig ab6966, with the DyLight 594 conjugated anti rabbit antibody ab96921. I would expect this to provide a good double staining. Please check however with your microscope specialist as well, whether this is an option for you. Excited at 543nm: DyLight® 550 : max excitation at 562nm, max emission at 576 (please see link below for more info on the spectrum) -goat anti rabbit IgG H+L, preabsorbed: ab96900 Click here (or use the following: https://www.abcam.com/Goat-Rabbit-IgG-HL-DyLight-550-preadsorbed-ab96900.html). Cy3.5: max excitation at 576nm, max emission at 589 -goat anti rabbit IgG H+L, preabsorbed: ab97076 Click here (or use the following: https://www.abcam.com/Goat-Rabbit-IgG-HL-Cy35--preadsorbed-ab97076.html). -goat anti guinea pig IgG H+L, preabsorbed: ab6966 Click here (or use the following: https://www.abcam.com/Goat-Guinea-pig-IgG-HL-Cy35-preadsorbed-ab6966.html). Texas Red: max excitiation at 595, max emission at 613 -donkey anti rabbit IgG H+L, preabsorbed: ab7081 Click here (or use the following: https://www.abcam.com/Donkey-Rabbit-IgG-HL-Texas-Red-preadsorbed-ab7081.html). TRITC: max excitiation at 557, max emission at 576 -goat anti guinea pig IgG H+L, preabsorbed: ab7136 Click here (or use the following: https://www.abcam.com/Goat-Guinea-pig-IgG-HL-TRITC-preadsorbed-ab7136.html). -donkey anti rabbit IgG H+L, preabsorbed: ab7080 Click here (or use the following: https://www.abcam.com/Donkey-Rabbit-IgG-HL-TRITC-preadsorbed-ab7080.html). Excited also 543nm: Cy3: max excitiation at 548, max emission at 561 -goat anti rabbit IgG H+L, preabsorbed: ab97075 Click here (or use the following: https://www.abcam.com/Goat-Rabbit-IgG-HL-Cy3-preadsorbed-ab97075.html). DyLight594: max excitiation at 591 max emission at 616 -we do have the anti guinea pig antibody in this conjugation, unfortunately not preabsorved. It would therefore need to be tested first. It is the ab96961. We might can provide you with a testing offer if you would be interested in this option. -donkey anti rabbit IgG H+L, preabsorbed: ab96921 Click here (or use the following: https://www.abcam.com/Donkey-Rabbit-IgG-HL-DyLight-594-preadsorbed-ab96921.html). As an alternative to an already conjugated antibody, I can also recommend to use either a biotynilated antibody or an unconjugated antibody. In those cases, you would either need an avidin/streptavidin in the right color or a correct conjugation kits (e.g. PerCP, tandem dyes). For the conjugation kits, I can recommend our easylink conjugation kits, very fast and easy to use: www.abcam.com/easylink Please do not hesitate to contact can we help or advise you further. We do have also a poster "guide to fluorochromes", which I can recommend to consult or to download: https://docs.abcam.com/pdf/immunology/fluorochrome_guide.pdf Alternatively, we do also have a poster on the DyLight antibodies. https://docs.abcam.com/pdf/general/DyLight_flyer.pdf I hope this information is helpful. Many season greetings!

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