PRODUCT CODE: ET7110-49

SIRT1 Recombinant Rabbit Monoclonal Antibody [JE11-04] (ET7110-49)

  • Recombinant

Applications

  • WB

  • ICC

REACTIVITY

  • Human

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+
Western blot analysis of SIRT1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-49, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Jurkat cell lysate<br />
Lane 2: HepG2 cell lysate<br />
Lane 3: Hela cell lysate
  • Western blot analysis of SIRT1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-49, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Jurkat cell lysate<br />
Lane 2: HepG2 cell lysate<br />
Lane 3: Hela cell lysate
  • ICC staining of SIRT1 in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7110-49, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Western blot analysis of SIRT1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-49, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Jurkat cell lysate
Lane 2: HepG2 cell lysate
Lane 3: Hela cell lysate

Applications

  • WB

  • ICC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

SIRT1 Recombinant Rabbit Monoclonal Antibody [JE11-04] (ET7110-49)

Immunogen

Synthetic peptide within c terminal human sirt1.

Host

Rabbit

Positive Control

Jurkat cell lysate, HepG2 cell lysate, Hela cell lysate, Hela.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JE11-04

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

Predicted band size 82 kDa.

Isotype

IgG

APPLICATION DILUTION

  • WB:1:500-1:2,000

  • ICC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

SIRT1

SYNONYMS

75SirT1 antibody; hSIR2 antibody; hSIRT1 antibody; HST2 antibody; HST2, S. cerevisiae, homolog of antibody; NAD dependent deacetylase sirtuin 1 antibody; NAD dependent protein deacetylase sirtuin 1 antibody; NAD-dependent deacetylase sirtuin-1 antibody; OTTHUMP00000198111 antibody; OTTHUMP00000198112 antibody; Regulatory protein SIR2 homolog 1 antibody; SIR1_HUMAN antibody; SIR2 antibody; SIR2 like 1 antibody; SIR2 like protein 1 antibody; SIR2, S.cerevisiae, homolog-like 1 antibody; SIR2-like protein 1 antibody; SIR2ALPHA antibody; SIR2L1 antibody; Sirt1 antibody; SirtT1 75 kDa fragment antibody; Sirtuin (silent mating type information regulation 2 homolog) 1 (S. cerevisiae) antibody; Sirtuin 1 antibody; Sirtuin type 1 antibody

SEQUENCE SIMILARITIES

Belongs to the sirtuin family. Class I subfamily.

TISSUE SPECIFICITY

Widely expressed.

POST-TRANSLATIONAL MODIFICATION

Methylated on multiple lysine residues; methylation is enhanced after DNA damage and is dispensable for deacetylase activity toward p53/TP53.; Phosphorylated. Phosphorylated by STK4/MST1, resulting in inhibition of SIRT1-mediated p53/TP53 deacetylation. Phosphorylation by MAPK8/JNK1 at Ser-27, Ser-47, and Thr-530 leads to increased nuclear localization and enzymatic activity. Phosphorylation at Thr-530 by DYRK1A and DYRK3 activates deacetylase activity and promotes cell survival. Phosphorylation by mammalian target of rapamycin complex 1 (mTORC1) at Ser-47 inhibits deacetylation activity. Phosphorylated by CaMK2, leading to increased p53/TP53 and NF-kappa-B p65/RELA deacetylation activity (By similarity). Phosphorylation at Ser-27 implicating MAPK9 is linked to protein stability. There is some ambiguity for some phosphosites: Ser-159/Ser-162 and Thr-544/Ser-545.; Proteolytically cleaved by cathepsin B upon TNF-alpha treatment to yield catalytic inactive but stable SirtT1 75 kDa fragment (75SirT1).; S-nitrosylated by GAPDH, leading to inhibit the NAD-dependent protein deacetylase activity.; Acetylated at various Lys residues. Deacetylated via an autocatalytic mechanism. Autodeacetylation at Lys-238 promotes its protein deacetylase activity.

SUBCELLULAR LOCATION

PML body, nucleus, cytoplasm, mitochondrion.

FUNCTION

The silent information regulator (SIR2) family of genes are highly-conserved from prokaryotes to eukaryotes and are involved in diverse processes, including transcriptional regulation, cell cycle progression, DNA-damage repair and aging. In S. cerevisiae, Sir2p deacetylates histones in an NAD-dependent manner, which regulates silencing at the telomeric, rDNA and silent mating-type loci. Sir2p is the founding member of a large family, designated sirtuins, which contain a conserved catalytic domain. The human homologs, which include SIRT1-7, are divided into four main branches: SIRT1-3 are class I, SIRT4 is class II, SIRT5 is class III and SIRT6-7 are class IV. SIRT1 has the closest homology to the yeast Sir2p and is widely expressed in fetal and adult tissues. SIRT1 is highly expressed in heart, brain and skeletal muscle, with low expression in lung and placenta. SIRT1 regulates the p53-dependent DNA damage response pathway by binding to and deacetylating p53, specifically at Lys 382.