Rabbit anti sirt3

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-SIRT3 is a primary antibody that recognizes the SIRT3 protein. SIRT3 is a member of the sirtuin family of NAD-dependent protein deacetylases. It is a mitochondrial protein that plays a role in regulating cellular metabolism and function.

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Lab products found in correlation

Image lab software, by Bio-Rad (2 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Complete mini tablet, by Roche (1 mentions) Mouse anti β actin antibody, by Santa Cruz Biotechnology (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Ultracruz mounting medium with dapi, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 488 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Mouse anti β tubulin, by Abcam (1 mentions) Rabbit anti sirt1, by Cell Signaling Technology (1 mentions) Horseradish peroxidase conjugated anti rabbit or anti mouse igg, by Merck Group (1 mentions) Mouse anti gapdh, by Cell Signaling Technology (1 mentions) Protease inhibitor cocktail, by Cell Signaling Technology (1 mentions) Enhanced chemiluminescence reagent, by Merck Group (1 mentions) Rabbit anti rac1, by Proteintech (1 mentions) Rabbit anti vegf, by Cell Signaling Technology (1 mentions) Rabbit anti hif 1α, by Cell Signaling Technology (1 mentions) Ripa buffer, by Merck Group (1 mentions)

Close spelling variants of this product

Rabbit SIRT3 Anti-SIRT3 SIRT3

4 protocols using rabbit anti sirt3

Glucose-Induced SIRT3 Expression in HRECs

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HRECs were exposed to glucose and transfected with miRNA mimics for 48 hr. Total proteins were prepared with RIPA buffer (Thermo Fisher) containing protease inhibitors (complete Mini Tablet; Roche). Protein concentrations were determined using Pierce™ BCA Protein Assay Kit (Thermo Scientific). A total of 30 μg of protein was resolved using SDS‐PAGE and transferred onto the PVDF membrane (Bio‐Rad), which was followed by blocking with 5% nonfat dry milk powder in TBS‐T and incubation with one of the primary antibodies: rabbit anti‐SIRT3 (1:1,000, Cell Signaling technology), mouse.
(AKL5C1) (1:1000, Santa Cruz) and mouse anti‐β‐actin antibody (1:1000, Santa Cruz) at 4°C overnight. After washing with TBS‐T, the membrane was incubated with HRP‐tagged secondary anti‐rabbit and anti‐mouse antibody (1:1000) for 1 hr at room temperature. Blots were visualized using electrochemiluminescence (Amersham Pharmacia Biotechnology). Relative band intensities were quantified by densitometry analysis using Image Lab software (Bio‐Rad).

Liu J., Chen S., Biswas S., Nagrani N., Chu Y., Chakrabarti S, & Feng B. (2020). Glucose‐induced oxidative stress and accelerated aging in endothelial cells are mediated by the depletion of mitochondrial SIRTs. Physiological Reports, 8(3), e14331.

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Immunofluorescent Analysis of SIRT3 and COXIV

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After deparaffinization, kidney tissue sections were treated with 10 mM citrate buffer (pH 6.0) for antigen retrieval. After blocking with 10% normal goat serum and 0.1% BSA at RT for one hour, the sections were incubated with rabbit anti-SIRT3 (1:300; Cell Signaling Technology Cat# 5490, RRID:AB_10828246) and mouse anti-COXIV (1:500; Santa Cruz Biotechnology) at 4°C overnight. The sections were then incubated with Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:500; Invitrogen, Carlsbad, CA, United States) and Alexa Fluor 550-conjugated goat anti-mouse IgG (1:500; Invitrogen, Carlsbad, CA, United States) at RT for one hour. After washing with PBS, slides were prepared and mounted using the UltraCruz™ Mounting Medium with DAPI (Santa Cruz Biotechnology, Santa Cruz, CA, United States) to detect nuclei. Images were captured on a Leica fluorescent microscope (Leica DM IRB, Germany) using a 20X/0.4 PH objective at 1.5-fold magnification, and the images were analyzed by the NIH ImageJ software (version 1.51).

Suliman H., Ma Q., Zhang Z., Ren J., Morris B.T., Crowley S.D., Ulloa L, & Privratsky J.R. (2021). Annexin A1 Tripeptide Mimetic Increases Sirtuin-3 and Augments Mitochondrial Function to Limit Ischemic Kidney Injury. Frontiers in Physiology, 12, 683098.

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Quantitative Western Blotting of Sirtuins

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Proteins were subjected to quantitative Western blotting as previously described [10, 20] using either rabbit-anti-SirT1 (1:1000, Cell Signalling Technology, Danvers, Massachusetts, USA, #2310), rabbit-anti-SirT2 (1:1000, Cell Signalling Technology, #2313), rabbit-anti-SirT3 (1:1000, Cell Signalling Technology, #2627), mouse-anti-NDUFA6 (0.1 µg/mL, Life Technologies, Paisley, UK, A31856) or anti-acetylated lysine monoclonal antibody (1:1000, Cell Signalling Technology, #9861), combined with horseradish peroxidase-conjugated antirabbit or anti-mouse IgG as appropriate (1:2000, Sigma). To normalise for protein concentration, membranes were stripped using Restore™ Western Stripping Reagent (Fisher Scientific, Loughborough, UK) and re-probed using a combination of mouse-anti-β-tubulin (1:2000, Abcam, ab7792) and rabbit-anti-mouse IgG, horseradish peroxidase conjugate (1:5000, Sigma). Images were digitally captured, band intensities quantified via densitometric analysis (with the exception of anti-acetylated lysine) using ImageLab software (BioRad, Hemel Hempstead, UK) and normalised for β-tubulin expression. Results were expressed as percentage expression compared to that observed in SH-SY5Y cells ± S.D (n = 4 for SirTs, n = 3 for NDUFA6).

Liu K.Y., Mistry R.J., Aguirre C.A., Fasouli E.S., Thomas M.G., Klamt F., Ramsden D.B, & Parsons R.B. (2015). Nicotinamide N-methyltransferase increases complex I activity in SH-SY5Y cells via sirtuin 3. Biochemical and biophysical research communications, 467(3).

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Protein Expression Analysis in Cell Extracts

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Total protein was extracted from cells were RIPA buffer (Sigma-Aldrich) containing Protease Inhibitor Cocktail (Cell Signaling Technology, Inc., Beverly, MA, USA). Protein concentration was determined using the bicinchoninic acid (BCA) protein assay kit (Thermo sher, Carlsbad, CA, USA). Equal amounts of protein extracts (40 µg) were separated by 10% SDS-PAGE and transferred to polyvinylidene di uoride membranes (Bia-Rad). Then the membranes were blocked and incubated overnight with rabbit anti-HIF-1α (Cell Signaling Technology), rabbit anti-SIRT3 (Cell Signaling Technology), rabbit anti-SDF-1α (Cell Signaling Technology), rabbit anti-VEGF (Cell Signaling Technology), rabbit anti-Rac1 (Proteintech Group.
Inc, Rosemont, IL, USA), rabbit anti-Rac2 (Proteintech Group. Inc), and mouse anti-GAPDH (Cell Signaling Technology) antibodies, respectively. The expression levels of protein were measured by enhanced chemiluminescence reagents (Millipore, Plano, TX, USA).

Chen X., Sun Y., Lei X., Cao Y., Gao M., Chen J., Bao B., & Chen L. (2021). HIF-1α overexpression in adipose mesenchymal stem cell-derived exosomes ameliorate hypoxia-induced dysfunction and inflammation in HUVECs.

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