Anti-Nestin抗体[10C2] - Neural Stem Cell Marker (ab22035)

Thank you for contacting us.

1. Anti-Nestin antibody [10C2] (ab22035) and Anti-HIF1 alpha antibody [H1alpha67] - BSA and Azide free (ab80544):

Volume: 100 µg at 1 mg/ml

We recommend to start with a concentration of 5 ug/ml. This will need further optimisation depending on samples and protocol.

2. Anti-c-Myc antibody [Y69] (ab32072)

Volume: 100 ul with a concentration from 0.1120 mg/ml for lot GR85451-5 (please refer to this lot when ordering).

We recommend to use this antibody in a dilution of 1/50 in IHC-P. As said previously this recommendation will need optimisation for every experiment by user.

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 very much for your interest in anti-Nestin antibodies.

To our knowledge our anti-Nestin products have not been tested in bovine. Therefore, I can offer a discount off a future purchase if you buy an anti-Nestin antibody now, test it in bovine for ICC and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free PRIMARY ANTIBODY.

While we do have 18 products against this target, I would suggest the following. These are either well characterized in the application or have been used in a number of species. I ordered them in what I feel is the most appropriate first but I can offer this discount for any or all of these.

ab6142: https://www.abcam.com/Nestin-antibody-2Q178-Neural-Stem-Cell-Marker-ab6142.html
ab27952: https://www.abcam.com/Nestin-antibody-Azide-free-ab27952.html
ab22035: https://www.abcam.com/Nestin-antibody-10C2-ab22035.html
ab81755: https://www.abcam.com/Nestin-antibody-ab81755.html

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test one or more of these antibodies in ICC on bovine. I will then send a discount code. This code must be issued before purchase so please wait for my reply before ordering.

2. Purchase either by phone, fax, or online (www.abcam.com).

3. Test it in bovine.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the antibody turns out to be unsuitable for bovine, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Thank you very much for your email. I apologize for the delay in our answer.

I can confirm thatthis antibody can be used with HRP conjugated secondary antibody without any problems.

The note on the datasheet is in reference on how it has been tested: the ab22035 has been tested on paraffin embedded tissue slides witha secondary antibody conjugated to FITC or Rhodamine. Inthat case, adilution of the ab22035 at1/100 to 1/200 gave good signal.

I would expect therefore, that using this antibody at 1/100 to 1/200 will also give good results using a secondary antibody coupled to HRP or the EXPOSE kit.

I hope this information is helpful. Please do not hesitate to contact us againshould you need any further information.

Thank you for your patience. I have attached the test results from the lab. We performed an immunoblotting experiment on a cell extract of U251 cells. The attached Western blot image clearly indicates a Nestin band with a MW of approximately 250 kDa. The test was a result of a 1:1000 antibody dilution prepared from a purified stock solution of 1mg/ml. The protein might be run higher than theoretically predicted due to the large number of phosphorylation sites (about 30). This however would not explain why you see a band at only 90 kDa. Please let me know if you have tried anything else in the meantime and receivd different results. If not and the antibody was ordered within the last 6 month, we can discuss a replacement, credit or refund. I hope this information is nevertheless helpful to you. Please do not hesitate to contact us if you need any more advice or information.