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

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

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.

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.

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.

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!

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!