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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 ab49931 for help.
Alternatively, you can search the previous enquiries about this product to see if your query has already been answered.
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DESCRIPTION OF THE PROBLEM No signal or weak signal
SAMPLE whole cell lysates(C2C12)
PRIMARY ANTIBODY Concentration or dilution - 1:2000 / 1:1000 Diluent buffer - 1% skim milk Incubation time - Over-night Incubation temperature: - 4oC Wash Buffer - TBS-T Number of washes - 3 min, 5times
DETECTION METHOD ECL (Amersham) & Super signal west Femto (Thermo scientific)
POSITIVE AND NEGATIVE CONTROLS USED -
ANTIBODY STORAGE CONDITIONS -20C
SAMPLE PREPARATION Lysis buffer :RIPA buffer Protease inhibitors: added Phosphatase inhibitors added
AMOUNT OF PROTEIN LOADED 15ug/lane
ELECTROPHORESIS/GEL CONDITIONS Percentage of gel - 12% SDS-PAGE
TRANSFER AND BLOCKING CONDITIONS Type of membrane - Nitrocellulose membrane Protein transfer verified -Ponceau S solution (sigma) Blocking agent and concentration - 5% skim milk Blocking time - 1hr~2hr Blocking temperature - Room temperature
SECONDARY ANTIBODY Goat anti-mouse IgG-HRP (SC-2005, santacruz) Concentration or dilution - 1:3000 dilution Diluent buffer - 1% skim milk Incubation time - 1hr Incubation temperature: - Room temperature Wash Buffer - TBS-T Number of washes - 3 min, 5times
HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes
WHAT STEPS HAVE YOU ALTERED? primary antibody dilution 1:2000 --> 1:1000
ADDITIONAL NOTES Our customer is keep using this antibody since 2008. This lot# showed too weak signal than before so wants to sned their data to technical team. |
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ANSWER: |
Thank you for confirming these details and for your cooperation.
I am sorry this batch seems to have a lower affinity than the other batches before have had. As the customer has used this antibody on mouse successfully before, we will replace the vial received with a new vial. Please ask the customer to submit an Abreview with his data so we do know that the clone normally works on mouse (as opposed to predicted to mouse as our current knowledge was).
Please confirm the customer would like to receive a new vial of a new lot of the ab49931.
I am looking forward to your reply. |
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BATCH NUMBER 582467 ORDER NUMBER 498 DESCRIPTION OF THE PROBLEM Non-specific staining SAMPLE C2C12 ( mouse myoblast cell line) PRIMARY ANTIBODY 1:50 in PBS, 1h binding, 3times wash DETECTION METHOD Conforcal microscopy POSITIVE AND NEGATIVE CONTROLS USED Positive control ( stabilin-2) ANTIBODY STORAGE CONDITIONS -20℃ FIXATION OF SAMPLE 4% paraformaldehyde ANTIGEN RETRIEVAL none PERMEABILIZATION STEP 0.3% Tritionx-100 BLOCKING CONDITIONS 4% BSA in PBS SECONDARY ANTIBODY 1:100 in PBS 30min, binding, 2times wash HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes |
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ANSWER: |
Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab49931 in ICC/IF. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support but please bear in mind that this product has not been tested in mouse species, only predicted to react, therefore it might not be suitable for use. I have looked through the protocols you used and it seems fine to me. Besides, you were able to obtain satisfactory results with a different antibody by using the same protocol. Therefore, I can only assume that in this occasion, this ab49931 is not suited for use in mouse species and we will not be able to offer our Abpromise guarantee. In any case, can I encourage you to submit an Abreview via the online product datasheet? We always encourage customers to send their results back to us, whether positive or negative, and we make all product information available to other researchers. We reward each Abreview with Abpoints which can be used for discounts off future purchases and gifts. If you decide to submit an Abreview, please include a note that you have already spoken to me, so that I will be able to quickly publish your Abreview, as I have already gone through your protocols. To find out more about our Abreview system, please see the following URL: http://www.abcam.com/abreviews Should you require any further information or assistance, please do not hesitate to contact me. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"