兔抗小鼠IgG H&L (HRP) (ab97046)

Thank you for your phone call. I did some research regarding PVA and found that for unknown reasons, PVA does not seem to effectively block background in WB. You may need to switch to BSA or milk in order to get clear bands in addition to the new secondary I am sending.
As requested, I have issued a free of charge replacement with the order number 1203530.

Regarding ab24070, the datasheet does indicate that B mercaptoethanol destroys the epitope, so you will need to be careful to avoid this reagent when blotting for Mucin 5AC.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Thank you for contacting us. Looking at your blots I think there is a few things you can try. One is to decrease the secondary antibody dilution. Since the blots are all the same it indicates some nonspecific binding. You can also add a no primary control to see if there is non-sepcific binding from the secondary alone. You can also increase your blocking time (even overnight at 4 degrees) or switch your blocking agent from milk to BSA or vice versa to see if this reduces background. Normally the primary antibody dilution for a WB when using a purified antibody can be about 1 ug/ml, however since the pattern is the same for all three different antibodies I am not convinced this is a primary antibody problem. Please let me know if these suggestions don't improve your results.

Thank you for contacting us.
The ab97023 and ab97046 are the antibodies from our AbExcel range; they are extensively tested, can be routinely used at dilutions ranging from 1/2000 to 1/20,000 and based on a 1/2000 dilution in 3ml milk/BSA, you can do over 600 blots with a single vial of HRP-conjugated AbExcel secondary. These are the reasons, why I recommended these products.
We have not performed western blot with IP lysates so we are unsure about the specificity towards native or denatured chains.
I can reassure you that these products are top of the range products and comes with full Abpromise guarantee. If because of any reason you are not satisfied with results please let us know we will help you with troubleshooting. Refund or credit note can also be given if you ended up being not happy at all with results.

I hope this information is helpful to you. We will look forward to receiving your order soon.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Thank you for contacting us.
We have better antibodies than ebioscience and these are stringently tested to get best results and best value per blot. Please click the following link for datasheet;
https://www.abcam.com/rabbit-polyclonal-secondary-antibody-to-mouse-igg-h-l-hrp-ab97046.html
https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-HL-HRP-ab97023.html
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Thank you for contacting us.
As this antibody has only been purified against it immunogen, it may cross- react with other species.
I have picked out three pre-adsorbed secondary antibodies, which show minimal cross-reactivity to human IgG:
https://www.abcam.com/Rabbit-Mouse-IgG-HL-HRP-preadsorbed-ab7075.html https://www.abcam.com/Rabbit-Mouse-IgG-HL-HRP-preadsorbed-ab7075.html.
https://www.abcam.com/Rabbit-Fab2-Mouse-IgG-HL-HRP-pre-adsorbed-ab98780.html https://www.abcam.com/Rabbit-Fab2-Mouse-IgG-HL-HRP-pre-adsorbed-ab98780.html.
https://www.abcam.com/Goat-Mouse-IgG-HL-HRP-preadsorbed-ab98808.html https://www.abcam.com/Goat-Mouse-IgG-HL-HRP-preadsorbed-ab98808.html.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Thank you for contacting us. I am sorry to hear these antibodies are not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. 1) About the secondary antibody ab6728: as indicated on the datasheet, many of our customers have reported seeing brown precipitates in the vials. The brown precipitates are very common with HRP conjugated antibodies; we suggest vortexing the vial and using this antibody as normal. However, I would be pleased to replace, if you wish, this antibody with an alternative catalog reference : ab97046, www.abcam.com/ab97046. 2) About the anti-His tag ab5000: to our knowledge this antibody has never been tested for one hour incubation only but overnight at 4ºC. Some antibodies bind better to their target at 4ºC, some other at room temperature. I think it would be worth trying an overnight incubation at 4ºC for this antibody. The blocking solution should not be problem, this antibody works fine in milk as well as in BSA. Please let me know about the results after an overnight incubation at 4ºC. If the antibody is still not working I will be pleased to offer a replacement or a credit note. Thank you for your cooperation. I look forward to hearing from you with your results using ab5000 and with details of how you would like to proceed with ab6728.

Thank you for contacting Abcam.
I can confirm that ab97406 is conjugated to HRP.
If there is anything else I can help you with, please let me know.