Net043

Manufactured by PerkinElmer

NET043 is a compact and versatile fluorescence detection system designed for a wide range of applications. It features high-performance optics and a sensitive detector to provide accurate and reliable fluorescence measurements.

Automatically generated - may contain errors

Lab products found in correlation

Amplify namp100, by Cytiva (1 mentions) Spin x centrifuge tube filter, by Corning (1 mentions) P9767, by Merck Group (1 mentions)

3 protocols using net043

Radiolabeled Fatty Acid Incorporation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bacteria grown for 2 days on SB plates were collected and resuspended in PBS to an OD₆₀₀ of 0.1. Six μl of bacterial suspension (approx. 3 × 10⁵ bacteria) were seeded in 3 ml of DMEM (41965-039; Gibco) containing 10% heat-inactivated human serum (HIHS) in 12-well plates (665 180; Greiner Bio-one). After 18 h of incubation at 37 °C with 5% CO₂, [9,10-³H] palmitic acid (32 Ci/mmol; NET043; Perkin-Elmer Life Sciences) was added to a final concentration of 50 μCi/ml and incubation was continued for 6 h. Bacteria were then collected by centrifugation, washed 2 times with 1 ml PBS and pellets were stored at −20 °C until further use. Pellets were treated with proteinase K as described above. Samples were loaded on 12% SDS PAGE gels and after electrophoresis, gels were fixed in 25:65:10 isopropanol:water:acetic acid solution overnight and subsequently soaked for 30 min in Amplify (NAMP100; Amersham) solution. Gels were vacuum dried and exposed to SuperRX autoradiography film (Fuji) for 12–18 hours until desired signal strength was reached.

Renzi F., Ittig S.J., Sadovskaya I., Hess E., Lauber F., Dol M., Shin H., Mally M., Fiechter C., Sauder U., Chami M, & Cornelis G.R. (2016). Evidence for a LOS and a capsular polysaccharide in Capnocytophaga canimorsus. Scientific Reports, 6, 38914.

+ Open protocol

+ Expand

Measuring Fatty Acid β-Oxidation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were labeled with 20 μCi/mL [9,10-³H]-palmitic acid (Perkin Elmer NET043) for 8 hrs at 37 °C. Tracer was removed by washing cells three times in PBS, and cells were serum starved in unlabeled medium for 16 hrs. For longer-term treatment, inhibitors were added for the entire 16-hr duration of the chase period. For shorter-term treatment, following the chase period, cells were pretreated for 1 hr with DGAT1 and 2 inhibitors or etomoxir prior to addition of torin1 for the indicated durations. Medium was refreshed for the final hour of the experiment to quantify production of ³H₂O, indicative of fatty acid β-oxidation. Medium was pelleted for 10 min at maximum speed at room temperature, and 350 μL were applied to 350 mg 1x8 Dowex Chloride Resin (Sigma 44340) that had been prewashed with water and dispensed into Spin-X Centrifuge Tube Filters (Costar). Tubes were centrifuged 3 min at 4000 x g at room temperature. ³H₂O in the flow through was quantified using Emulsifier Safe scintillation cocktail on a Hidex 300 SL counter. Cells were lysed in NP-40 lysis buffer and protein was quantified to normalize ³H₂O produced.

Hosios A.M., Wilkinson M., McNamara M.C., Kalafut K.C., Torrence M.E., Asara J.M, & Manning B.D. (2022). mTORC1 regulates a lysosome-dependent adaptive shift in intracellular lipid species. Nature metabolism, 4(12), 1792-1811.

+ Open protocol

+ Expand

Palmitate Uptake in Soleus Muscle

Check if the same lab product or an alternative is used in the 5 most similar protocols

Soleus muscle was removed from mice of each genotype and immediately soaked in DMEM supplemented with 0.5 μCi/mL [9,10‐³H]‐palmitate (PerkinElmer, NET043‐1000 μCi) in the presence of 2% BSA and 0.2 mM cold palmitate (Sigma, P9767). After 30 min incubation at 30°C, tissues were washed thoroughly and lysed with 50°C 1 M KOH solution for 1 h. Samples were then used for scintillation counting.

Kim A., Koo J.H., Jin X., Kim W., Park S., Park S., Rhee E.P., Choi C.S, & Kim S.G. (2021). Ablation of USP21 in skeletal muscle promotes oxidative fibre phenotype, inhibiting obesity and type 2 diabetes. Journal of Cachexia, Sarcopenia and Muscle, 12(6), 1669-1689.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!