I am interested in a source of antibodies against proteins encoded by mouse mitochondrial DNA. I am aware that some companies advertise such. Therefore, my interest is in those that were validated in your lab. Ideally, on WT vs. rho-0 lysates.
Some of my co-workers validated the COX I (MS604 from mitosciences, host specie mouse) in rat samples.
Which of Mitosciences or Invitrogen's antibodies were validated in your lab? Can you provide catalog numbers? We tested most, if not all Mitosciences' antibodies with the only bright spot-COXI antibody mentioned by Susana (although, MS604 is catalog number for a mix of 5 antibodies, COXI (ab14745(MS304)) being only one of them.
Am interested in any Antibodies against any Proteins encoded by mtDNA or nDNA. Have you or your team published any papers on this subject - human, mouse or any ? Could defects in mtDNA or nDNA (encoded mt Proteins) directly or as a consequence stimulate/trigger antibodies to proteins outside the mitochondria or dependent on OXPHOS function etc - leading to antibodies against important cellular proteins causing disruption, inflammation or consequential disease or pseudodisease of speiific organs (eg. thyroid A/Bs, pituitary A/Bs, pancreatic A/Bs ?
Hi Mikhail, I found your question because I was looking for the same info :)
I wonder whether you were able to find these antibodies?
COX sub-unit I antibodies are being used to indicate mitochondrial protein synthesis. I am looking for another method to indicate the same thing for verification purposes, like other antibodies for example. I have not checked on mito-RNA, but this could be an indicator for mtDNA translation activity
What do you guys think??
So far, no luck. With one exception, all antibodies we tested failed WB test on wt vs rho-0 lysates. You can observe mitochondrial protein synthesis by inhibiting cytosolic protein synthesis with emetine and doing metabolic labeling with 35S met/cys with/without chloramphenicol as mitochondrial protein synthesis inhibitor.
sounds a good idea .. Is it a known method for this propose? Where can I find more details/reference
Hi, I'm looking to do a similar experiment to test the mitochondrial translation in human cells. Any advice for human mtDNA encoded protein antibodies I could use. Looking for recommendations.
Did any of you try the mtND antibodies (e.g. Abcam and SCBT)?
Back in the day, we tested all abcam's antibodies (and you may see some of our reviews on their website). One exception that works is the mtCO1 antibody. The caveat is, it ONLY works with non-denatured samples (found that out through tech's sloppiness). Another caveat is that we are looking for MOUSE reactivity (some, but not all, of abcam's antibodies DO work with human lysates). The third caveat is that these are polyclonals. Some dramatic lot-to-lot variations are possible, esp. with attentition being paid to QC. We did not retest recent lots (it has been 5 yrs since the original post). As far as SC is concerned, they have only one mtnd antibody, human-specific (therefore, of little interest to us).
Annie, ab14745 from abcam recognizes both human and mouse mtCOI. You should not boil your samples, though. There were some other human-specific antibodies from abcam. I believe, some of their mtCO2 worked quite well. I can look that up
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