Anti-alpha Tubulin antibody - Loading Control (ab4074)

Phone call requesting refund for non-specificity of ab4074 and reporting dissatisfaction with alteration in volume of antibody provided.

ANSWER:

Thank you for calling us yesterday to report your concerns over the specificity of theanti-alpha tubulin antibody (ab4074) as well as the changes in our vial sizes you have noticed.

As discussed over the phone, from time totime we do need to change the sizes we provide of our products. This is not a universal strategy in order to make more money from our products but due to a number of different reasons, often outside of our own control. For example, sometimes an antibody which was being produced in good scale becomes more difficult to produce and/or characteriseand we therefore need to reduce the quantity provided as it no longer becomes financially viable to provide it in the same size. We constantly review such factors and try our best to take customeropinion into consideration. Therefore, if you could provide me with a list of the antibodies you are no longer satisfied with due to the change in size provided I would be grateful. I can then pass this on to the relevant people within Abcam to look into why these changes have been made and if we can do anything to amend the change.

As discussed over the phone, I have also sent out a newvial of theanti-KIF17antibody(ab11261) to you. This should be with you today. In order to understand your concerns further, could you briefly explain how you have been using this antibody? Are you again performing ICC? In a 1/100 dilution? I have also arranged for the order of anti-alpha tubulin (ab4074) to be refunded.

I am going to contact the lab over how the anti-ensconsin fastrack antibody (ab79124) is being dealt with. I have looked through your correspondence with my colleague Miia about this antibody and understand your concerns over its specificity. I will get back to you as soon as I have further information relating to this.

I look forward to receiving your reply.

Two images are attached in a powerpoint file. The protocol as I said on the phone is standard for the lab - we've used it for years in this and other cell lines on a wide variety of cytoskeletal proteins, including other microtubule antibodies. If necessary, I can provide images of microtubules stained with another rabbit primary in this cell line.
Cells: Human TC7 colon cancer cell line.
Protocol:
Fixed in methanol (5 mins @ -20)
Blocked in Goat Serum (10%, 30 mins)
Dilution 1/50, for 1 hour at room temperature
Secondaries: Goat anti-rabbit alexaflour 488 (first image) and 647 (second image), 30 mins at room temperature, dilution 1/1000

ANSWER:

Thank you very much for your email.

I was also in discussion with lab regarding the specificity of this antibody. They confirmed that they have seen some cross reactivity with junction like proteins in ICC/IF however only when they use methanol for fixation. This antibody provided good results with PFA which is why the lot was stocked for sales so they recommend using PFA as fixative.

I hope this info will also be helpful.

Two images are attached in a powerpoint file. The protocol as I said on the phone is standard for the lab - we've used it for years in this and other cell lines on a wide variety of cytoskeletal proteins, including other microtubule antibodies. If necessary, I can provide images of microtubules stained with another rabbit primary in this cell line.
Cells: Human TC7 colon cancer cell line.
Protocol:
Fixed in methanol (5 mins @ -20)
Blocked in Goat Serum (10%, 30 mins)
Dilution 1/50, for 1 hour at room temperature
Secondaries: Goat anti-rabbit alexaflour 488 (first image) and 647 (second image), 30 mins at room temperature, dilution 1/1000

ANSWER:

Thank you for your email.

I have checked the images you have kindly sent and agree that this antibody seems like binding to cell junction proteins. I have sent the image to lab and now having discussion with them regarding the specificity of this lot. Lab may retest this lot however it not decided yet; once they decide to retest it then it is going to take further 2 weeks to receive the results.

This time line may affect your progression in the project so I am happy to provide a vial of different lot as a replacement antibody to avoid any inconveniance.

Could you let me know what you think?

ab4074 binding non specifically to cell junction proteins.
- Used at 1/50 dilutions
- The secondary antibody and protocol is fine because the same protocol was successfully with other antibodies.

ANSWER:

Thank you very much for your phone call. I am sorry to hear that you are experiencing problems using this antibody.

I have checked the history of this antibody so I can say that there are not many complaints we have received for this ab so as we discussed I am happy to discuss the image with lab. Could you please send the images and brief protocol you have used?

Thank you! I will look forward to hearing from you soon.

Can this ab be used against bacterial samples?

ANSWER:

Thank you very much for your interest in ab4074. I am have not been able to determine definatelty if one way or another, if there is a good chance that ab4074 would work for your sampletypes. Therefore, I can offer a discount off a future purchase if you buy ab4074 now, test it in E.Coli 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. 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 ab4074 in E.Coli. I will then send a discount code. This code must be issued before purchasing ab4074 so please wait for my reply before ordering. 2. Purchase ab4074 either by phone, fax, or online (www.abcam.com). 3. Test it in E.Coli. 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: http://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 ab4074 turns out to be unsuitable for E.Coli, 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.