You have changed your country from to
. Please be aware that this will change the currency in the purchasing process.
Related products
- Secondary antibodies
- Lysates
Reassurance, Refunds & Replacements
If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »This product is covered by the Abpromise guarantee. Our scientific support team are available to answer any questions or queries - fill out an inquiry form for ab13793 for help.
Alternatively, you can search the previous enquiries about this product to see if your query has already been answered.
|
|||||||||
 | |||||||||
|
|||||||||
Blocking with 10 % nonfat dry milk in TBST 1 hour at RT. Primary antibody diluted in 3% nonfat dry milk in TBST |
|||||||||
ANSWER: |
Thank you very much for answering my questions. I'm sorry to hear you are having a problem with ab13793. I would like to suggest the following modifications to your protocol: Please try blocking with only 5% milk in TBST. 10% seems high to me, you may be blocking the membrane too thoroughly. Also, please try removing the milk from the primary antibody solution, or using a low percentage such as 0.5%. Sometimes the blocking agent in the primary solution prohibits the antibody from binding. Please let me know if this helps and do not hesitate to contact us for further advice. |
||||||||
|
|||||||||
BATCH NUMBER 75947 ORDER NUMBER 64848 DESCRIPTION OF THE PROBLEM No band at all SAMPLE K562 cell extract PRIMARY ANTIBODY anti-MBD4 (ab13793), 1/100 and 1/200 dilution in TBST, overnight 10 min three time wash with TBST DETECTION METHOD ECL and ECF both ANTIBODY STORAGE CONDITIONS stored in the fridge SAMPLE PREPARATION Laemmli buffer AMOUNT OF PROTEIN LOADED 100 micro gram ELECTROPHORESIS/GEL CONDITIONS reducing, 4-15 % gradient gel TRANSFER AND BLOCKING CONDITIONS regular transfer buffer with 1.5 hr SECONDARY ANTIBODY [competitor], Alka-phos conjugated and HRP conjugated HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? increasing primary antbody concentation changing secondary antibody detection method. ADDITIONAL NOTES I used other primary antibody ( at least 9 different types) in the same gel and blot everytime. I got specific bands or nonspecific backgraound with other primary, but not with anti-MBD4 antibody. I purchased anti-DNA pol beta antibody at the same time, it worked everytime. It is unlikely that rabbit polyclonal antibody does not work at 1/100 dilution.
|
|||||||||
ANSWER: |
Thank you for your enquiry. Could you please tell me how you blocked the membrane? Also, did you add blocking agent to your primary antibody? I hope this information helps, please do not hesitate to contact us if you need any more advice or information. |
||||||||
|
|||||||||
| |||||||||
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
