Protocol for preparing mitochondrial fraction for native PAGE?

07 July 2018 0 4K Report

I have isolated mitochondria from the tissue of a tapeworm and want to perform NADH fumerate reductase activity staining using native PAGE process. How should I proceed?

Badges
Science method

More Bidisha Ukil's questions See All

NADH oxidase and Succinate dehydrogenase specific activity and percentage inhibition calculation?

I have performed NADH oxidase and succinate dehydrogenase assay from mitochondrial fraction from the cestode parasite Hymenolepis diminuta which undergoes anaerobic energy metabilism. I want to...

04 May 2018 6,891 2 View

Calculation procedures for estimating protein carbonyls in tissue samples?

protein carbonyls are oxidative stress markers. after following a specific procedure for estimating the change in the activity of protein carbonyls the process of calculating their concentration...

08 September 2017 1,986 4 View

Do I need specific antibodies to stain for a fusion protein?

I am staining some brain sections stored in cryoprotectant that express a Histone H2B- GFP fusion protein that were generated ~10 years ago. I know I need to enhance signal with an anti-GFP...

07 August 2024 5,338 2 View

Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

How to determine positive-stained cells in FACS? Use isotype or unstained control?

To compare positive and negative cell populations in flow cytometry, should I compare unstained cells with antibody stained cells? Or with the isotype control? Most papers show comparison with...

06 August 2024 6,728 6 View

Can I multiplex a mouse monoclonal and rat primary for IHC (mouse brain tissue)?

Hi all, I was just wondering if anyone has experience with multiplexing a mouse monoclonal primary and a rat primary. I'm trying to multiplex by incubating them in the same well but was told by a...

06 August 2024 9,710 1 View

How to preform densitometry on SDS-page bands?

I ran a SDS-page of a bacterial lysate and I want to quantify protein concentration in a specific band. I was thinking of using a standards ladder or make some standards are different...

05 August 2024 9,805 3 View

Senescence-associated beta galactosidase staining is False Positive in control group?

Hi guys If anyone is currently working on aging cells, you guys would like to give me some advice. I'm testing against biomarker (SA-beta-Gal), I encountered a false positive in the control group...

02 August 2024 6,735 1 View

Following click reaction in cell lysates, protein is immobile and remains at the top of the gel in SDS-PAGE?

I am using CuBr/THPTA for a click reaction in total cell lysates. I am facing issues with my protein sample in non-reducing SDS-PAGE where it's not migrating properly and most of it remains at the...

29 July 2024 950 4 View

Suggestions for the Annexin V Positive Control ?

Hello everyone, I am currently using washed human platelets to stain Annexin V as a procoagulant marker. Additionally, I am staining with PerCP-CD61 to identify platelet cells. So far, I have...

29 July 2024 8,624 1 View

How can I get clearer background for my western blot?

I am running a western on HIV envelope protein. The size is around 50Kda and I am using rat anti-HA for the first stain and a second stain of mouse anti-CA. Primary antibody dilution is 1: 1000...

29 July 2024 6,484 5 View

How many events should be counted for the analysis of apoptosis with nuclear staining with Hoechst 33342 whit microscopy ?

Greeting How many events should be counted for the analysis of apoptosis with nuclear staining with Hoechst 33342 whit microscopy ? Are 100 events count sufficient and three repetitions for...

25 July 2024 7,520 0 View