Anti-RIP140 抗体 (ab42126)

Thank you for contacting us.

I can confirm, that to our knowledge, customised protocols are not required for the use of ab42126 - Anti-RIP140 antibody - ChIP Grade in IP. However, I can recommend to have a look at our standard protocol, provided on our Abcam Homepage here:

https://www.abcam.com/index.html?pageconfig=resource&rid=11385

https://www.abcam.com/index.html?pageconfig=resource&rid=11382

Furthermore, I would suggest to review the provided references provided for the use of this antibody on it's datasheet.

1. Ho PC et al. NF-?B-mediated degradation of the coactivator RIP140 regulates inflammatory responses and contributes to endotoxin tolerance. Nat Immunol 13:379-86 (2012).

"Cells were then collected and lysed in SDS lysis buffer"

2. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3395431

I hope this information is helpful to you. Please do not hesitate to contact us if you have any problems or if you need any more advice or information.

Thank you for your inquiry and your patience with this reply. I apologize that it took the testing laboratory a while to confirm how this antibody has been further tested. The lab confirmed that the antibody has been tested for labeling endogenous expression of this protein in cellular extracts, but since the protein is mainly present in nuclear extracts, the extraction is sometimes difficult.  They recommended a strong lysis buffer, such as RIPA buffer, or even doing a nuclear extraction (protocol linked below). RIPA buffer (Radio Immuno Precipitation Assay buffer) 150 mM sodium chloride 1.0% NP-40 or Triton X-100 0.5% sodium deoxycholate 0.1% SDS (sodium dodecyl sulphate) 50 mM Tris, pH 8.0 https://www.abcam.com/index.html?pageconfig=resource&rid=11408 Also since this is such a large protein, they recommended using an 8% gel. In addition, large proteins will tend to precipitate in the gel, hindering transfer. Adding SDS to a final concentration of 0.1% in the transfer buffer will discourage this. Methanol tends to remove SDS from proteins, so reducing the methanol percentage to 10% or less will also guard against precipitation. Plus, lowering methanol in the transfer buffer also promotes swelling of the gel, allowing large proteins to transfer more easily. But methanol is only necessary if using nitrocellulose. If using PVDF, methanol can be removed from the transfer buffer altogether, and is only needed to activate the PVDF before assembling the gel/membrane sandwich. Also you should choose to do a wet transfer overnight at 4oC instead of semi-dry transfer. Lastly the primary antibody was incubated 1/1000 overnight at 4C. If these suggestions do not improve your results and you have purchased the antibody in the last 6 months, I would be happy to replace or refund it for you. I hope this information helps and please let me know if you have any other questions.

free of charge replacement with ab3425

Thank you for contacting us. This antibody was generate from an 18 aa peptide from the C terminal.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us.

This product's country of origin is the United States.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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