Sirtuin 2/SIRT2 Antibody [Biotin]

SIRT2 expression is increased in free fatty acid (FFA)-exposed tolerant RAW 264.7 cell macrophages (RAW) cells. Bovine serum albumin (BSA: vehicle for free fatty acid) or stearic acid (free fatty acid: FFA)-exposed RAW264.7 cell macrophages (RAW) were stimulated with either one or two doses of the LPS; second dose at indicated time points after 1st LPS. (A) TNF-alpha mRNA expression was evaluated by qRT–PCR analysis. (B) SIRT2 protein expression was detected by Western blot in BSA- or FFA-exposed RAW cells following a single dose of the LPS stimulation for indicated time periods. GAPDH was used as the loading control. (C) Western blot image quantification using image-J software (n = 3 each group; * p < 0.05). SIRT2 protein expression was normalized to GAPDH. (D,E). To inhibit SIRT2, after 4 h of 1st LPS exposure, cells were treated with SIRT2 inhibitor AK-7 (25 µM) and incubated further for 20 h in the presence or absence of the second dose of the LPS. TNF-alpha protein released in the cell culture media from FFA-exposed sensitive and tolerant cells was evaluated by ELISA. (* p < 0.05). IL6 (Figure 1E) protein released in the cell culture media from FFA-exposed sensitive and tolerant cells evaluated by ELISA. (* p < 0.05). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33810233), licensed under a CC-BY license. Not internally tested by R&D Systems.

The effect of acute ethanol-exposure-induced SIRT2 on PFKP expression in macrophages. (A) PFKP expression in WT-BMDM exposed to vehicle or ethanol ± LPS by western blot analysis. (B) PFKP western blot image quantification of PFKP protein in vehicle vs. ethanol exposed WT-BMDM, Y axis represents fold of vehicle-LPS (fold of vehicle control) (n = 4 blots; * p < 0.05). (C) PFKP expression in Ethanol-exposed WT-BMDM and SIRT2KO-BMDM ± LPS. (D) Western blot image quantification of PFKP protein in ethanol exposed WT-BMDM and SIRT2KO-BMDM. Y axis represents fold of Ethanol-exposed WT-LPS (fold of WT- ethanol control) (n = 4 blots; * p < 0.05). (E) PFKP expression in Ethanol-exposed WT-BMDM treated with AK-7 or DMSO ± LPS. (F) Western blot image quantification of PFKP in AK-7 vs. DMSO treated-Ethanol-exposed WT-BMDM ± LPS, Y axis represents fold of Ethanol-exposed WT-LPS (fold of WT- ethanol control) (n = 4 blots; * p < 0.05). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36865524), licensed under a CC-BY license. Not internally tested by R&D Systems.

The effect of acute ethanol-exposure on mouse bone marrow derived macrophages (BMDM). Phagocytosis in BMDM exposed to vehicle or ethanol ± LPS to study phagocytosis using pHrodo bioparticles and SIRT2 expression. (A) Representative images of intracellular pHrodo bioparticles in vehicle or Ethanol-exposed WT-BMDM ± LPS. (B) Fluorescence quantification of pHrodo bioparticles in WT-BMDM (n=4 repetitions/group; * p<0.05). (C) SIRT2 protein expression detected by western blot in Vehicle- or Ethanol-exposed BMDM cells ± LPS. (D) Western blot image quantification of SIRT2 protein blot in vehicle or ethanol-exposed BMDM cells ± LPS (n = 4 blots/group; * p < 0.05). (E) Representative images of pHrodo bioparticles in Ethanol-exposed WT-BMDM and SIRT2KO-BMDM ± LPS. (F) Fluorescence quantification of pHrodo bioparticles in Ethanol-exposed WT-BMDM, and SIRT2KO-BMDM ± LPS. * p < 0.05. (G) Representative images of pHrodo bioparticles in Ethanol-exposed WT-BMDM, co-treated with SIRT2 inhibitor AK-7 or DMSO ± LPS. (H) Fluorescence quantification of pHrodo bioparticles in Ethanol-exposed WT-BMDM with AK-7/DMSO ± LPS stimulation. * p < 0.05. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36865524), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effect of K394R mutation on PFKP. (A, B) HEK293T cells transfected with wtPFKP/mtPFKP in the presence or absence of SIRT2. Western blot analysis of Turbo-GFP-wtPFKP, DDK-SIRT2, turbo-GFP (control for wtPFKP plasmid transfected) and CPA. (C) mtPFKP transfection and IP using turbo-GFP-trap in HEK293T cells, in presence or absence of SIRT2 followed by IB analysis of acetyl lysine, turbo-GFP-mtPFKP and turbo-GFP (control for wtPFKP plasmid transfected). Pulldown with HEK293T cell lysate without transfection was used as a negative control. (D) Western blot analysis of turbo-GFP mtPFKP, DDK-SIRT2, turbo-GFP and CPA in whole cell lysate, used as an input for the turbo-GFP IP. (E) mtPFKP transfection and IP using magnetic-TUBEs in HEK293T cells, in presence or absence of SIRT2 followed by IB analysis of ubiquitination. (F) Western blot analysis of turbo-GFP mtPFKP, DDK-SIRT2, turbo-GFP and CPA in whole cell lysate used as input for the TUBE IP. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36865524), licensed under a CC-BY license. Not internally tested by R&D Systems.

The effect of acute ethanol-exposure-induced SIRT2 on PFKP expression in macrophages. (A) PFKP expression in WT-BMDM exposed to vehicle or ethanol ± LPS by western blot analysis. (B) PFKP western blot image quantification of PFKP protein in vehicle vs. ethanol exposed WT-BMDM, Y axis represents fold of vehicle-LPS (fold of vehicle control) (n = 4 blots; * p < 0.05). (C) PFKP expression in Ethanol-exposed WT-BMDM and SIRT2KO-BMDM ± LPS. (D) Western blot image quantification of PFKP protein in ethanol exposed WT-BMDM and SIRT2KO-BMDM. Y axis represents fold of Ethanol-exposed WT-LPS (fold of WT- ethanol control) (n = 4 blots; * p < 0.05). (E) PFKP expression in Ethanol-exposed WT-BMDM treated with AK-7 or DMSO ± LPS. (F) Western blot image quantification of PFKP in AK-7 vs. DMSO treated-Ethanol-exposed WT-BMDM ± LPS, Y axis represents fold of Ethanol-exposed WT-LPS (fold of WT- ethanol control) (n = 4 blots; * p < 0.05). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36865524), licensed under a CC-BY license. Not internally tested by R&D Systems.

The effect of acute ethanol-exposure on mouse bone marrow derived macrophages (BMDM). Phagocytosis in BMDM exposed to vehicle or ethanol ± LPS to study phagocytosis using pHrodo bioparticles and SIRT2 expression. (A) Representative images of intracellular pHrodo bioparticles in vehicle or Ethanol-exposed WT-BMDM ± LPS. (B) Fluorescence quantification of pHrodo bioparticles in WT-BMDM (n=4 repetitions/group; * p<0.05). (C) SIRT2 protein expression detected by western blot in Vehicle- or Ethanol-exposed BMDM cells ± LPS. (D) Western blot image quantification of SIRT2 protein blot in vehicle or ethanol-exposed BMDM cells ± LPS (n = 4 blots/group; * p < 0.05). (E) Representative images of pHrodo bioparticles in Ethanol-exposed WT-BMDM and SIRT2KO-BMDM ± LPS. (F) Fluorescence quantification of pHrodo bioparticles in Ethanol-exposed WT-BMDM, and SIRT2KO-BMDM ± LPS. * p < 0.05. (G) Representative images of pHrodo bioparticles in Ethanol-exposed WT-BMDM, co-treated with SIRT2 inhibitor AK-7 or DMSO ± LPS. (H) Fluorescence quantification of pHrodo bioparticles in Ethanol-exposed WT-BMDM with AK-7/DMSO ± LPS stimulation. * p < 0.05. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36865524), licensed under a CC-BY license. Not internally tested by R&D Systems.

Effect of K394R mutation on PFKP. (A, B) HEK293T cells transfected with wtPFKP/mtPFKP in the presence or absence of SIRT2. Western blot analysis of Turbo-GFP-wtPFKP, DDK-SIRT2, turbo-GFP (control for wtPFKP plasmid transfected) and CPA. (C) mtPFKP transfection and IP using turbo-GFP-trap in HEK293T cells, in presence or absence of SIRT2 followed by IB analysis of acetyl lysine, turbo-GFP-mtPFKP and turbo-GFP (control for wtPFKP plasmid transfected). Pulldown with HEK293T cell lysate without transfection was used as a negative control. (D) Western blot analysis of turbo-GFP mtPFKP, DDK-SIRT2, turbo-GFP and CPA in whole cell lysate, used as an input for the turbo-GFP IP. (E) mtPFKP transfection and IP using magnetic-TUBEs in HEK293T cells, in presence or absence of SIRT2 followed by IB analysis of ubiquitination. (F) Western blot analysis of turbo-GFP mtPFKP, DDK-SIRT2, turbo-GFP and CPA in whole cell lysate used as input for the TUBE IP. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/36865524), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Biotin
• Concentration: LYOPH

Sirtuin 2/SIRT2 Antibody [Biotin] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human Sirtuin 2/SIRT2
  Ala2-Gln389
  Accession # Q8IXJ6
• Specificity: Detects human Sirtuin 2/SIRT2 in Western blots. In Western blots, less than 1% cross-reactivity with recombinant human SIRT1 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: SIRT2
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 ug/mL
• Readout System:
  • Streptavidin Native Protein NBP2-29367
  • Streptavidin Native Protein NBP2-29370
  • Streptavidin Native Protein NBP2-29371

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Sirtuin 2/SIRT2 Antibody [Biotin]

• EC 3.5.1
• EC 3.5.1.-
• FLJ35621
• FLJ37491
• S.cerevisiae, homolog) 2
• SIR2L2
• SIRT2
• Sirtuin 2
• sirtuin type 2

Background

Sirtuin 2, encoded by the SIRT2 gene, is also known as SIR2 (silent information regulator 2)-like protein 2. It is a nicotinamide adenine dinucleotide (NAD)-dependent histone/protein deacetylase (1, 2). The SIR2 family of enzymes is classified as class III histone deacetylases (HDACs) and has been implicated in many cellular processes that include histone deacetylation, gene silencing, chromosomal stability, and aging (3, 4). Unlike class I and class II HDACs, the enzymatic activity of class III HDACs is NAD dependent and insensitive to HDAC inhibitor trichostatin A (5).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Sirtuin 2/SIRT2 Antibody (BAF4358)(2)

Publications using BAF4358

• Gandhirajan A, Roychowdhury S, Kibler C et al. SIRT2-PFKP interaction dysregulates phagocytosis in macrophages with acute ethanol-exposure Frontiers in Immunology 2023-01-27 [PMID: 36865524]
• Sanjoy Roychowdhury, Anugraha Gandhirajan, Christopher Kibler, Xianfeng Wang, Vidula Vachharajani Sirtuin 2 Dysregulates Autophagy in High-Fat-Exposed Immune-Tolerant Macrophages Cells 2021-03-26 [PMID: 33810233]

Bioinformatics

• Gene Symbol: SIRT2
• Uniprot:
  • Q8IXJ6()