Anti-Bcl2 antibody [100/D5] (ab692)

Hi,



I have some antibodies from you, and their datasheets say that I need to store them at 4C, for example, ab98711 and ab692.

However, most of antibodies from abcam need to be stored at –20C or –80C.



So, my question is what is the estimated shelf life of the product if I store it at 4C as its datasheet says.

I’m worried whether it is correct that they are loosing their affinity or denatured with a higher rate compared to other antibodies kept at –20C as time goes by.
And, what happens if I freeze it?

Thank you in advance.

ANSWER:

Thank you for your enquiry.

Estimated shelf life of antibodies stored at 4 degrees C is about 1 year, although frequently they last much longer.

I recommend that you follow the storage conditions as noted on the product datasheet. Freezing an antibody that is intended to only be stored at 4 degrees C may lead to denaturing of the antibody.

I hope this is helpful. Please contact me again if you have any further questions.

Hi.

I have frozen Ab692 by a mistake. The freezer's temperature is around
-30 degrees. What should I do? Should I take it out and put it in -80 or
transfer it to a refrigerator at 4 degrees? Do you think the antibody is
still usable for IHC?

Regards,

ANSWER:

Thank you for your enquiry. I am sorry to hear you have some concerns regarding the storage of the antibody.

I would suggest to take the antibody out of the freezer and put it at +4oC. The antibody solution does contain some BSA, which can act as a cryoprotectant, so the antibody may be ok and I would certainly recommend it is worth trying in an experiment. However, regrettably I am sorry we would not longer be able to guarantee how well this will work.

If it has lost some activity, you could try using it at a higher concentration.

I hope this will be helpful. Please do not hesitate to let me know if you have any further questions. Good luck with the experiments.

BATCH NUMBER 127954 ORDER NUMBER -- NOT SPECIFIED --

DESCRIPTION OF THE PROBLEM No staining

SAMPLE Human colon tissue

PRIMARY ANTIBODY Mouse monoclonal to Bcl2 : abcam 1:25-50 in PBS- 1% BSA At RT for 1hour or at 4? O/N incubation PBS 3min 2times washing

DETECTION METHOD ABC kit (peroxidase ) / DAB and ABC kit (alkaline phosphatase)/Fast blue

POSITIVE AND NEGATIVE CONTROLS USED Positive control ? colon cancer I detected P21, and p53 but could not detect Bcl2 using same sample.

ANTIBODY STORAGE CONDITIONS 4 ?

FIXATION OF SAMPLE Paraformaldehyde

ANTIGEN RETRIEVAL Heat mediated

PERMEABILIZATION STEP 0.01 M citrate buffer (pH 9.5)

BLOCKING CONDITIONS 10 % goat serum at RT for 30min

SECONDARY ANTIBODY biotinylated anti-mouse IgG(H+L) : vector BA-2000 1:300 in PBS- 1% BSA at RT for 30min PBS 3min 2times washing

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 10 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

WHAT STEPS HAVE YOU ALTERED? Ab dilution, incubation condition

ANSWER:

Thank you for your enquiry. I am sorry to hear that your customer has been having difficulties with this antiserum. This antiserum has received very few complaints.

I have read the questionnaire from your customer and I have a few comments. Please can you ask your customer to confirm that they have been using this antiserum against more than one colon cancer cell preparation. I am a little concerned that Bcl2 is not showing detectable levels of expression in the cells that you are using.

I would also be interested to know whether your customer has tried any permeabilization steps and/or changing the length of time for the antigen retrieval step. Bcl2 is an integral mitochondrial protein and therefore it may be that the relative accessibility of the antigen is the limiting step in the success of this antibody by IHC-P.

I hope this information helps.

Hi, I would like information about the anti bcl2 antibody (cat number ab692). I would like to get the price for it and also at which concentration should I use it for western blot. Also, is it possible to send me a picture of how a western blot should look like with this antibody.

Thank you very much

ANSWER:

Thank you for your email. Ab692 is $295 (US dollars). To use ab692 in Western blotting I would recommend starting at a concentration of 1 ug/ml and optimizing from there based on results. I'm afraid that we do not have a Western blot image available at this time.

If you have any additional questions, please contact us again.

Thank you, but a little too late. The antibody does not work for western. I used another from Calbiochem, which works perfectly. I found out that the concentration was 200ug/ml about a month ago by searching another company's catalog.

Not very good customer service on Abcam's part at all. You should know everything (at the very least the concentration) about the antibodies you are selling. I will no longer order from Abcam.

ANSWER:

Thank you for your feedback. I apologise for any inconvenience that you have suffered. We make every attempt to add as much detail to our product data sheets as possible. We produce some of our antibodies in-house and others are sourced from suppliers whose products we believe will be of interest to our customers. This product falls into the latter category. Western blotting was added as an additional application in attempt to provide further data to the customer the concentration of the product unfortunately took time to confirm. I am happy to offer you a full refund to the value of this product, please could you confirm your order details to allow me to proceed with our accounts department. Once again, my apologies. I look forward to hearing from you soon.