Bovine Heart Mitochondria (ab110338)

Submit a review Q&A (22)References (3)

Overview

  • Product name

    Bovine Heart Mitochondria

  • General notes


    2 mg of bovine heart mitochondria in iso-osmotic buffer. This sample can act as a positive control in immunocapture applications and Blue Native PAGE.

    1 mg solubilized mitochondria for Immunocapture experiments. For the Immunocapture of OXPHOS complexes, it is recommended that 180 µl of the mitochondrial membranes supplied be solubilized by the addition of 20 µl of 10% lauryl maltoside. Insoluble material is removed by centrifugation at 72,000g before immunocapture of complexes from the supernatant. For more detail see Abcam's suggested protocols.

  • Tested applications

    Suitable for: IP, SDS-PAGE, WBmore details

Properties

  • Mycoplasma free

    Yes

  • Form

    Liquid

  • Storage instructions

    Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.

  • Storage buffer

    Constituents: 0.02% PMSF, 0.16% Tris, 0.89% EDTA, 8.56% Sucrose
  • Concentration information loading...

  • Lysate notes

    2 mg of bovine heart mitochondria in iso-osmotic buffer. This sample can act as a positive control in immunocapture applications and Blue Native PAGE.

    1 mg solubilized mitochondria for Immunocapture experiments. For the Immunocapture of OXPHOS complexes, it is recommended that 180 µl of the mitochondrial membranes supplied be solubilized by the addition of 20 µl of 10% lauryl maltoside. Insoluble material is removed by centrifugation at 72,000g before immunocapture of complexes from the supernatant. For more detail see Abcam's suggested protocols.

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab110338 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP
Use at an assay dependent concentration.
SDS-PAGE
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
Notes
IP
Use at an assay dependent concentration.
SDS-PAGE
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (3)

Publishing research using ab110338? Please let us know so that we can cite the reference in this datasheet.

ab110338 has been referenced in 3 publications.

  • Andreux PA  et al. Mitochondrial function is impaired in the skeletal muscle of pre-frail elderly. Sci Rep 8:8548 (2018). PubMed: 29867098
  • Gómez-Serrano M  et al. Differential proteomic and oxidative profiles unveil dysfunctional protein import to adipocyte mitochondria in obesity-associated aging and diabetes. Redox Biol 11:415-428 (2017). Functional Studies ; Human . PubMed: 28064117
  • Shell JR & Lawrence DS Proteolytic regulation of the mitochondrial cAMP-dependent protein kinase. Biochemistry 51:2258-64 (2012). PubMed: 22385295

Customer reviews and Q&As

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11-20 of 22 Abreviews or Q&A

Question

I am working in a research department, so I don’t track the lot#. I mentioned earlier I don’t know whether three runs are with same lot, last run is GR71915 on the box. I attached the packing list when I received the package. Looks like there are three different lots. I used last two kits today which one from GR82148-1, and one is GR82148-2.

I will try heating samples on 37C and with the lysate ab when I receive it. By the way, what is 2nd ab for this lysate ab, is it the same in the kit?

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Answer

Thank you for contacting us.

The lysate I am sending you free of charge is just that - a lysate (ab110338, Bovine Heart Mitochondrial lysate, https://www.abcam.com/bovine-heart-mitochondrial-lysate-ab110338.html)

You would load it onto your gel like your other samples, and then use the antibodies in the kit - as you previously did to detect the proteins in the membrane after transfer from the gel.

You would treat the bovine heart mito lysate the same way you treat your samples (e.g. add SDS loading buffer, warm up to 37 degree, add DTT freshly).

Please let me know what your results look like.

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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Question

Thank you for your e-mail.

I just found out today with my 3rd western as you discovered, the week band is COX-1 protein. I had western two years ago, signal from COX-1 protein is much more strong then the high unknown band. I attached some pictures, one from two years ago and recent data for your investigation. Three runs from recent data were using three different kits. I really concern why that unknown band is much stronger comparing two years ago, whether this will alter our results/data.

When I run the western, the sample buffer has fresh DTT, no heating. Please let me know any root cause for high unknown band.

By the way, please send me the two replacement kits.

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Answer

Thank you for your email.

I have issued a free of charge replacement with the order number xxx for 2 kits with new lot GR82148-3 (for the kit component issue).
I have also arranged for a goodwill shipment of the bovine heart mitochondrial lysate ab110338. This is on order number xxx - free of charge - for trying the WB with this lysate in comparison with your samples.


Regarding your data: We do not know what the higher MW band could be.
We recommend heating to 37 degree. If you do not heat at all, the denaturing process might not be as efficient as when done with some amount of heat.

Is there a way to find out which lot numbers you have tried in the 3 recent attempts? It would be important to know for us if the lotnumbers are the same or different. You had mentioned you used lot GR82148-1 in the recent tries. Is the third attempt from the same or another lot?

Please let me know what results you obtain with the positive control lysate that I am sending you you.

I look forward to hear back.


Regarding the free of charge shipments: To check the status of the orders 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.

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Question

I’m very interested in your pyruvate dehydrogenase activity assay kit (ab109902), but I’m concerned about the samples I’d like to test. They are rat heart tissue pieces that were flash frozen in liquid nitrogen and stored at -80… for about 6 years. If I were only quantifying the protein I wouldn’t be overly concerned, but I’d like to assay activity. I have successfully isolated RNA for real-time PCR, and the RNA seems fine.   If you could advise me on whether these samples are probably still ok to test for activity, I would greatly appreciate it.

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Answer

Thank you for contacting us. If the samples were properly stored at -80C without thawing/freezing cycles, we feel think that the PDH activity will be preserved. If there is enough material, we would advise you to isolate mitochondria from a portion of one of the samples and test both the whole tissue and also the mitochondrial prep. We also recommend a good positive control such as ab110338,Bovine Heart Mitochondrial lysate. https://www.abcam.com/Bovine-Heart-Mitochondrial-lysate-ab110338.html We are also currently running a 4 for 3 promotion. With purchase of any 3 Abcam products you will receive a 4th absolutely free. If you are interested in this please using this code when ordering: xxxxx 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

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Question



Thanks for your reply. Our lab is not allowed to send details of our clinical protocols, so unfortunately we can't do that. We do however do these assays routinely, and since they are working with other control samples, we feel it is a vial or batch issue, and nothing to do with our protocol.

We did speak to Abcam and he said that we would be sent a replacement vial to see if it was just an isolated problem. We said we would get back to him with feedback once we tried it out.

I will consult with our secretary to get the Abcam confirmation number and PO number from her, but Tom had managed to find the order when I spoke to him on the phone. If you could check that a replacement vial has been sent to us that would be great, as he had implied that he was going to do it, and we do need to rectify this problem as soon as possible.


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Answer

Thank you for your response. It has been arranged for this product to be delivered to you. The order confirmation number is XXXX. This product is also covered by our Abpromise guarantee. If should continue to experience difficulties using this product within six months, we will replace or refund it under our Abpromise guarantee, as you are using it according to specifications listed on our datasheet. I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Question

The MitoScience Beef Heart mitochondria ( Cat # MS802) that we received on Feb 3, 2011 had a lot # D2066. I used it until  April this year when I received abCAM , Cat #AB110338, Lot# GR48373-20.

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Answer

Thank you for sending these additional details.

I am sorry that you have been experiencing difficulties using this product. Both lotsD2066 and GR48373-20 are taken from the same stock so it seems possible that the trouble which you have been experiencing is a vial specific issue. If you could please send me details about your protocol for our internal investigations as well as the Abcam order confirmation number or PO used I would be happy to provide a free of charge replacement for you.

I look forward to your reply. Please let me know if you have any questions.

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Question

We have been using bovine Heart Mitochondria from MitoSciences (Cat # MS802) as a control for measuring respiratory enzymes in muscle biopsies. Our last purchase came from abCAM (Lot# GR48373-20) and the rates for some of the enzymes are different. Could it be just this particular lot# or has anything been added to the mitochondrial pellet that might have an effect on the mitochondrial enzymes? As I have already mentioned we use your product only as a control for measuring mitochondrial enzymes so we could generate new ranges if your protocol for mitochondrial isolation has changed, as long as we know about the change.

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Answer

Thank you for contacting Abcam.

Would you happen to have the lot numbers of the previous aliquots you have used?

I look forward to you reply. Please let me know if you have any questions or there are other ways we may help you.

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Question

I am seeing much less mitochondrial complex I and III activity from the most recent vial of this lysate. Has production changed at all since 2008?

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Answer

The MitoSciences lab sent the following information regarding the bovine heart mitochondria today:

"In regards to the retail lots in question (E1405 and D2066), they are made from the same stock lot of concentrated, purified BHM (lot D0707, 33 mg/mL).

The only difference then between D2066 and E1405 is that D2066 was thawed and aliquoted directly into the retail vial by MitoSciences (this lot was before we integrated with Abcam), whereas E1405 was thawed and aliquoted into a bulk vial, which was then thawed again by Abcam into retail vials. This extra freeze/thaw cycle could potentially account for the difference in enzyme activity."

(Lot D2066 is the one you received in 2008, and lot E1405 is the more recent one, and is the only lot we have in stock now).

The MitoSciences lab also pointed out:

"We do not normally QC MS802 (ab110338), but we happened to do a Western blot QC test of lot E1405 because it was part of our integration stocking order. I have attached the data. You’ll see on the blot that there are strong bands for the each of the representative subunits of Complex I and Complex III, but of course expression does not necessarily translate into activity."

Please let me know if you would like to see the blot, and do not hesitate to contact us if you need any other information or if you have any concerns.

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Question

I would like to dilute this further, but do not have any information about the buffer it is in. How should I dilute this for my WB?

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Answer

Thank you for contacting Abcam regarding ab110338.

I have confirmed that the mitochondria are in an iso-osmotic buffer.
For dilution in Western blot the sample can be diluted in SDS-PAGE WB buffer.

I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

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Question

are bovine heart mitochondria (not as lysate) still being available (used to beMS802)?

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Answer

Thank you for contacting us.

I have checked with the lab:

For ab110338, the used term "lysate" is unfortunately incorrect.The buffer constituents indicatethat the mitochondria are in an iso-osmotic buffer, not a lysis buffer. The labhastriedto get the term 'lysate' removed from these products for a while asthis is misleading. I will look into this.

But to answer you question: Essentially ab110338 isyou arelooking for - these arethe actual mitochondria.

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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Question

uses human skeletal muscle biopsies from children (i.e. has limited samples)
How much tissue do I need to startwith?
Is this different for each kit?

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Answer

Thank you for contacting us.

I have heard back from the lab with the following information:

All these assays require different sample concentrations described in each booklet protocol - but in most cases is approximately 10ug/microplate well.
The good news is that normal whole muscle homogenate contains a good robust mitochondria signal so less is required, muscle samples are described in most if not all protocols.
Another good news is that the same preparation for any of the assays is suitable for all others. I would estimate that if a 1 mg of homogenate can be prepared (0.2mL at 5mg/mL).
This would provide enough material for each of the assays when diluted to the specific concentration in each assay.
We would recommend - if contemplating doing multiple assays - to include a mitochondrial sample as positive control such as ab110338 isolated mitochondria (note this is cow source)



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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11-20 of 22 Abreviews or Q&A

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