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Sirt3

Manufactured by Thermo Fisher Scientific
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SIRT3 is a laboratory equipment product designed for use in scientific research. It functions as a measurement tool for the analysis of the Sirtuin 3 protein, which is involved in cellular metabolism and energy production. The core function of SIRT3 is to enable researchers to quantify and study the activity of this specific protein in various experimental settings.

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Lab products found in correlation

Abc kit, by Vector Laboratories (1 mentions) Confocal microscopy, by Zeiss (1 mentions) Biotinylated goat anti rat igg, by Vector Laboratories (1 mentions) Goat anti mouse texas red conjugated igg, by Merck Group (1 mentions) Foxo3, by Proteintech (1 mentions) Sirt1, by Thermo Fisher Scientific (1 mentions) Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions) Gapdh, by Thermo Fisher Scientific (1 mentions) Vinculin, by Thermo Fisher Scientific (1 mentions) Histone h3, by Thermo Fisher Scientific (1 mentions) Cardiac troponin t, by Thermo Fisher Scientific (1 mentions) Cox 4, by Bio-Rad (1 mentions) Actinβ, by Thermo Fisher Scientific (1 mentions) Acetonitrile, by Merck Group (1 mentions) Acetic anhydride, by Merck Group (1 mentions) Zeba spin desalting column, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

SIRT3 siRNA (4 citations) Anti-Sirt3 (2 citations)

4 protocols using sirt3

1

Immunohistochemical and Immunofluorescence Analysis of SIRT3 and FOXO3

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Tissues fixed with 4% formalin were embedded in paraffin. Then, tissue sections were incubated with primary antibody of SIRT3 (Invitrogen, Carlsbad, CA, USA) or FOXO3 (Proteintech, Wuhan, China). Biotinylated goat antirat IgG (Vector, Burlingame, CA, USA), as the secondary antibody, was incubated with immunoperoxidase (ABC Kit, Vector) following the manufacturer’s protocols. SIRT3 and FOXO3 in BMDMs were detected by immunofluorescence with antirabbit SIRT3 pAb (Invitrogen, Carlsbad, CA, USA) and antimouse FOXO3 mAb (Proteintech, Wuhan, China), followed by incubation with secondary goat antirabbit Texas Green-conjugated IgG or goat antimouse Texas Red-conjugated IgG (Sigma, St. Louis, MO, USA). DAPI was used to stain the nuclei. The slides were washed twice with PBS and detected using confocal microscopy (ZEISS, Oberkochen, Germany) in accordance to the manufacturer’s protocols. Positive cells were observed and counted in 10 HPF/section (×200 or ×400).
Wang Q., Wei S., Li L., Qiu J., Zhou S., Shi C., Shi Y., Zhou H, & Lu L. (2020). TGR5 deficiency aggravates hepatic ischemic/reperfusion injury via inhibiting SIRT3/FOXO3/HIF-1ɑ pathway. Cell Death Discovery, 6, 116.
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2

Long-term siRNA-mediated gene silencing

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The PSilencer 3.1-H1 linear vector from Ambion Inc. (Austin, TX) was used to obtain long term gene silencing. To target mouse Nfkbie, a siRNA Oligo Duplex (Locus ID 18037) from Origen (Rockville, MD) was used. The siRNAs to target mouse Nfe2l2 (RefSeq NM_010902.3), Ppargc1 (RefSeq NM_008904.2), Sirt1 (RefSeq NM_019812.2), Sirt3 (RefSeq MN_022433.2), Parp1 (RefSeq NM_007415.2), SOD2 (RefSeq NM_013671.3), GPX1 (RefSeq NM_008160.6), and PRDX2 (peroxiredoxin 2) (RefSeq NM_009116.2) were from Invitrogen (San Diego, CA). In all cases silencing procedures followed the technical recommendations of the manufacturers. In the control experiments we used equivalent amounts of the corresponding sense oligonucleotides and scrambled oligonucleotides with the same base composition and a randomized sequence. Silencing was confirmed by immunoblotting. Invitrogen Lipofectamine RNAiMAX Transfection Reagent and manufactureŕs protocol were used for delivery of siRNA.
Obrador E., Salvador-Palmer R., Pellicer B., López-Blanch R., Sirerol J.A., Villaescusa J.I., Montoro A., Dellinger R.W, & Estrela J.M. (2022). Combination of natural polyphenols with a precursor of NAD+ and a TLR2/6 ligand lipopeptide protects mice against lethal γ radiation. Journal of Advanced Research, 45, 73-86.
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3

Antibody Preparation for Immunoblotting

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Antibodies used for immunoblots were purchased from the indicated companies: MST1 (BD-611052 and H00006789-M02), GAPDH (CS-2118), P-LATS (CS-8654), LATS (SC-398560), Histone H3 (CS-9715), cl-CAS3 (CS-9661), cl-CAS9 (CS-9509), Actin-β (SC-69879), SIRT3 (SC-365175 and CS-5490), Vinculin (sc-25336), H2B (CS-12364), Pser14-H2B(CS-6959), Tri-Methyl-Histone (Lys27) H3 (CS-9733), Cardiac Troponin T (Thermo MA5-12,960), COX IV (CS- 4850) [BD = Bio-Rad; CS = Cell Signaling; H = Invitrogen; SC = Santa Cruz]. Anti-mouse and anti-rabbit HRP-linked antibodies were purchased from Bio-Rad and Cell Signaling Technology. All antibodies were diluted in 5% non-fat dry milk powder in 0.05% Tween 20 Tris-buffered saline (TBST) or 3% BSA in TBST.
Schirone L., Vecchio D., Valenti V., Forte M., Relucenti M., Angelini A., Zaglia T., Schiavon S., D’Ambrosio L., Sarto G., Stanzione R., Mangione E., Miglietta S., Di Bona A., Fedrigo M., Ghigo A., Versaci F., Petrozza V., Marchitti S., Rubattu S., Volpe M., Sadoshima J., Frati L., Frati G, & Sciarretta S. (2023). MST1 mediates doxorubicin-induced cardiomyopathy by SIRT3 downregulation. Cellular and Molecular Life Sciences, 80(9), 245.
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4

Acetylation and Deacetylation of Cartilage Homogenate

Cited 1 time
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Cartilage total homogenate was prepared in phosphate buffer as described for glutathione measurements. Cartilage homogenate was acetylated in vitro using 25μM acetic anhydride (Sigma-Aldrich, 539996) in acetonitrile (Sigma-Aldrich, 271004) for 2 min at room temperature (23 (link)). Cartilage homogenate was deacetylated in vitro using 1μg recombinant human SIRT3 (Enzo Life Science International, BML-SE270-0500) in the presence or absence of 1mM NAD+ and then incubated at room temperature for 5 minutes following previously described methods (24 ). After SIRT3 treatment, samples were desalted through Zeba™ Spin Desalting Column (Thermo Scientific, 89882) to remove dithiothreitol (DTT). Desalted samples were stored on ice prior to conducting SOD2 activity assays. Although NAD+ catalyzes the activity of other sirtuin isoforms that are present at endogenous levels in the cartilage homogenate, only SIRT3 interacts specifically with SOD2 (25 (link)).
Fu Y., Kinter M., Hudson J., Humphries K.M., Lane R.S., White J.R., Hakim M., Pan Y., Verdin E, & Griffin T.M. (2016). Aging Promotes SIRT3-dependent Cartilage SOD2 Acetylation and Osteoarthritis. Arthritis & rheumatology (Hoboken, N.J.), 68(8), 1887-1898.
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