Human Sirtuin 1/SIRT1 Antibody

Catalog #: MAB7714 Datasheet

Catalog #	Availability	Size / Price	Qty
MAB7714
MAB7714-SP

Sirtuin 1/SIRT1 in HepG2 Human Cell Line.

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Human Sirtuin 1/SIRT1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Sirtuin 1/SIRT1 in ELISAs. In direct ELISAs, no cross-reactivity with recombinant human Sirtuin 2, 3, 5, or 6 is observed.

Source

Monoclonal Mouse IgG₁ Clone # 834918

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human Sirtuin 1/SIRT1
Met1-Ser747
Accession # Q96EB6

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL

See below

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunocytochemistry

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Sirtuin 1/SIRT1 in HepG2 Human Cell Line. Sirtuin 1/SIRT1 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Mouse Anti-Human Sirtuin 1/SIRT1 Monoclonal Antibody (Catalog # MAB7714) at 25 µg/mL for 1 hour at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Intracellular Staining by Flow Cytometry

Detection of Sirtuin 1/SIRT1 antibody in HepG2 Human Cell Line antibody by Flow Cytometry.

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Detection of Sirtuin 1/SIRT1 in HepG2 Human Cell Line by Flow Cytometry. HepG2 human hepatocellular carcinoma cell line was stained with Mouse Anti-Human Sirtuin 1/SIRT1 Monoclonal Antibody (Catalog # MAB7714, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Background: Sirtuin 1/SIRT1

SIRT1 (SIR2-like protein 1; also NAD-dependent protein deacetylase sirtuin-1 and hSIR2) is a class I member of the sirtuin family of enzymes. Although its predicted MW is 81 kDa, it runs anomalously at 110-120 kDa in SDS-PAGE. It is a widely expressed nuclear protein that participates in the deacetylation of multiple proteins, including p300, p53, LKB1 and histone H1. Functionally, this has the effect of promoting heterochromatin formation, cell survival and resistance to oxidative stress. Metabolically, SIRT1 induces insulin secretion, inhibits glycolysis and suppresses fatty acid synthesis. Human SIRT1 is 747 amino acids (aa) in length. It possesses two NLSs (aa 32-39 and 223-230), an NES (aa 138-145), and a sirtuin-type deacetylase domain (aa 241-495) that contains an NAD and Zn binding motif. There are at least 12 utilized Ser/Thr phosphorylation sites, plus two nitrosylated Cys and one acetylated Ala. There are also four potential isoform variants. One is 95 kDa in size and shows a deletion of aa 454-639, a second is 17 kDa in size and contains a 16 aa substitution for aa 149-747, and a third contains an alternative start site at Met296. SIRT1 is also known to undergo proteolysis by cathepsin B at Val533Ser534, generating a fourth, C-terminally truncated 75 kDa isoform. Full-length SIRT1 is suggested to form trimers, while the 17 kDa isoform appears to form dimers. Over aa 2-747, human and mouse SIRT1 share 86% aa sequence identity.

Long Name

Silent Mating Type Information Regulation 2 Homolog

Entrez Gene IDs

23411 (Human); 93759 (Mouse); 309757 (Rat)

Alternate Names

EC 3.5.1; S. cerevisiae, homolog) 1; SIR2alpha; SIR2L1; SIR2-like protein 1; SIRT1; Sirtuin 1