Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, Western Blot, Flow Cytometry, CyTOF-ready

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all Sirtuin 1/SIRT1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Sirtuin 1/SIRT1
Ala2-Ser747
Accession # Q96EB6

Specificity

Detects human Sirtuin 1/SIRT1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human SIRT2 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human Sirtuin 1/SIRT1 Antibody

Detection of Human Sirtuin 1/SIRT1 antibody by Western Blot.

Detection of Human Sirtuin 1/SIRT1 by Western Blot.

Western blot shows lysates of A172 human glioblastoma cell line, A549 human lung carcinoma cell line, and HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human Sirtuin 1/SIRT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7714) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Sirtuin 1/SIRT1 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Sirtuin 1/SIRT1 antibody in HepG2 Human Cell Line antibody by Flow Cytometry.

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

HepG2 human hepatocellular carcinoma cell line was stained with Sheep Anti-Human Sirtuin 1/SIRT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7714, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). 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.
Western Blot Shows Human Sirtuin 1/SIRT1 Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human Sirtuin 1/SIRT1 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and SIRT1 knockout HeLa cell line (KO). PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human Sirtuin 1/SIRT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7714) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Sirtuin 1/SIRT1 at approximately 120 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

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.

Flow Cytometry

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

Knockout Validated

Sirtuin 1/SIRT1 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Sirtuin 1/SIRT1 knockout HeLa cell line.

Western Blot

0.2 µg/mL
Sample: A172 human glioblastoma cell line, A549 human lung carcinoma cell line, and HeLa human cervical epithelial carcinoma cell line

Reviewed Applications

Read 1 review rated 5 using AF7714 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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 NLS's (aa 32-39 and 223-230), an NES (aa 138-145), and a sertuin-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

Certificate of Analysis

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Product Specific Notices for Human Sirtuin 1/SIRT1 Antibody

For research use only

Citations for Human Sirtuin 1/SIRT1 Antibody

Customer Reviews for Human Sirtuin 1/SIRT1 Antibody (1)

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  • Human Sirtuin 1/SIRT1 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293 human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 01/06/2018
    Human Sirtuin 1/SIRT1 Antibody AF7714

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