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You recently helped me to find an HA-tagged protein (CARHSP1) that I could use for quantification on Western blots. I have just tested it out, and it does run and bind the anti-HA antibody nicely, but unfortunately there are 3 bands (approx 44, 35 and 30 kDa) instead of just the 44 kDa one. This will make it harder to use it for quantification but I think we will still try (using quantification software), as it's the only product available.
I wondered if you knew what these extra bands are, and whether they would be present in a particular proportion that we can pre-calculate? I expect some of the protein is just degraded. .....
Asked on Sep 05 2012
Thank you for your email and your patience.
The producers of this protein confirmed that these bands are degradation products. Unfortunately we do not have another batch without these bands.I am sorry for this inconvenience.
Please see here the comments from the originator on the image in the attachment andseen on ourdatasheet: "The three bands present on the gel are as follows – 44 kDa GST-HA-CRSPH24 (full length), 35 kDa and 30 kDa are both degraded forms for the GST-HA-CRSPH24. I have attached a copy of the SDS-Page gel for this protein and you can clearly see the three bands and it corresponds with the anti-HA western blot.We haven't calculated the appropriate proportion of the full length to the breakdown, but the full length is clearly the majority.Unfortunately we do not have a batch of the protein without these degradation fragments."
I hope you will be able to use this protein nevertheless to quantify your results. In order to apologize for the inconvenience, I can offer a 50% discount on the purchase ofa second unit of this protein. Please let me know if this would be an interesting option for you.
Thank you also for submitting an Abreview. We look forward to receive your feedback!
Thank you for your cooperation. Please do not hesitate to contact us, should you have any other question or concern.
Answered on Sep 05 2012