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.

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.

Applications for Human Sirtuin 1/SIRT1 Antibody

Application

Recommended Usage

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
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: HepG2 human hepatocellular carcinoma cell line were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

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.

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

Alternate Names

SIR2-like protein 1, SIR2alpha, SIR2L1, SIRT1

Entrez Gene IDs

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

Gene Symbol

SIRT1

UniProt

Q96EB6 (Human)

Additional Sirtuin 1/SIRT1 Products

Product Documents for Human Sirtuin 1/SIRT1 Antibody

Product Datasheets

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COA

SDS

Download SDS

Product Specific Notices for Human Sirtuin 1/SIRT1 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human Sirtuin 1/SIRT1 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB7714

Key Product Details

Species Reactivity

Validated:

Applications

Label

Antibody Source

View all Sirtuin 1/SIRT1 Antibodies »

Product Summary for Human Sirtuin 1/SIRT1 Antibody

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human Sirtuin 1/SIRT1 Antibody

Sirtuin 1/SIRT1 in HepG2 Human Cell Line.