Anti-SIRT1抗体[19A7AB4] (ab110304)

概述

  • 产品名称Anti-SIRT1抗体[19A7AB4]
    参阅全部 SIRT1 一抗
  • 描述
    小鼠单克隆抗体[19A7AB4] to SIRT1
  • 经测试应用适用于: IP, In-Cell ELISA, WB, Flow Cyt, ICC/IF, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Recombinant Human SIRT1

  • 阳性对照
    • HDFn and HL60 cells; HepG2, H9C2 and MEF cell lysates. Human normal colon FFPE tissue.

性能

应用

Our Abpromise guarantee covers the use of ab110304 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IP Use at an assay dependent concentration.
In-Cell ELISA Use a concentration of 4 µg/ml.
WB Use a concentration of 0.125 µg/ml. Predicted molecular weight: 81 kDa.

Detects a band of approximately 110 kDa (110-121 kDa) which is likely to be due to post translational glycosylation. SIRT1 is known to bind to several other proteins, and the 121 kDa band could also be due to the presence of one of these complexes (ensure samples are adequately reduced and denatured).

Flow Cyt Use a concentration of 1 µg/ml.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 0.5 µg/ml.
IHC-P Use a concentration of 5 µg/ml.

靶标

  • 功能NAD-dependent protein deacetylase that links transcriptional regulation directly to intracellular energetics and participates in the coordination of several separated cellular functions such as cell cycle, response to DNA damage, metobolism, apoptosis and autophagy. Can modulate chromatin function through deacetylation of histones and can promote alterations in the methylation of histones and DNA, leading to transcriptional repression. Deacetylates a broad range of transcription factors and coregulators, thereby regulating target gene expression positively and negatively. Serves as a sensor of the cytosolic ratio of NAD(+)/NADH which is altered by glucose deprivation and metabolic changes associated with caloric restriction. Is essential in skeletal muscle cell differentiation and in response to low nutrients mediates the inhibitory effect on skeletal myoblast differentiation which also involves 5'-AMP-activated protein kinase (AMPK) and nicotinamide phosphoribosyltransferase (NAMPT). Component of the eNoSC (energy-dependent nucleolar silencing) complex, a complex that mediates silencing of rDNA in response to intracellular energy status and acts by recruiting histone-modifying enzymes. The eNoSC complex is able to sense the energy status of cell: upon glucose starvation, elevation of NAD(+)/NADP(+) ratio activates SIRT1, leading to histone H3 deacetylation followed by dimethylation of H3 at 'Lys-9' (H3K9me2) by SUV39H1 and the formation of silent chromatin in the rDNA locus. Deacetylates 'Lys-266' of SUV39H1, leading to its activation. Inhibits skeletal muscle differentiation by deacetylating PCAF and MYOD1. Deacetylates H2A and 'Lys-26' of HIST1H1E. Deacetylates 'Lys-16' of histone H4 (in vitro). Involved in NR0B2/SHP corepression function through chromatin remodeling: Recruited to LRH1 target gene promoters by NR0B2/SHP thereby stimulating histone H3 and H4 deacetylation leading to transcriptional repression. Proposed to contribute to genomic integrity via positive regulation of telomere length; however, reports on localization to pericentromeric heterochromatin are conflicting. Proposed to play a role in constitutive heterochromatin (CH) formation and/or maintenance through regulation of the available pool of nuclear SUV39H1. Upon oxidative/metabolic stress decreases SUV39H1 degradation by inhibiting SUV39H1 polyubiquitination by MDM2. This increase in SUV39H1 levels enhances SUV39H1 turnover in CH, which in turn seems to accelerate renewal of the heterochromatin which correlates with greater genomic integrity during stress response. Deacetylates 'Lys-382' of p53/TP53 and impairs its ability to induce transcription-dependent proapoptotic program and modulate cell senescence. Deacetylates TAF1B and thereby represses rDNA transcription by the RNA polymerase I. Deacetylates MYC, promotes the association of MYC with MAX and decreases MYC stability leading to compromised transformational capability. Deacetylates FOXO3 in response to oxidative stress thereby increasing its ability to induce cell cycle arrest and resistance to oxidative stress but inhibiting FOXO3-mediated induction of apoptosis transcriptional activity; also leading to FOXO3 ubiquitination and protesomal degradation. Appears to have a similar effect on MLLT7/FOXO4 in regulation of transcriptional activity and apoptosis. Deacetylates DNMT1; thereby impairs DNMT1 methyltransferase-independent transcription repressor activity, modulates DNMT1 cell cycle regulatory function and DNMT1-mediated gene silencing. Deacetylates RELA/NF-kappa-B p65 thereby inhibiting its transactivating potential and augments apoptosis in response to TNF-alpha. Deacetylates HIF1A, KAT5/TIP60, RB1 and HIC1. Deacetylates FOXO1 resulting in its nuclear retention and enhancement of its transcriptional activity leading to increased gluconeogenesis in liver. Inhibits E2F1 transcriptional activity and apoptotic function, possibly by deacetylation. Involved in HES1- and HEY2-mediated transcriptional repression. In cooperation with MYCN seems to be involved in transcriptional repression of DUSP6/MAPK3 leading to MYCN stabilization by phosphorylation at 'Ser-62'. Deacetylates MEF2D. Required for antagonist-mediated transcription suppression of AR-dependent genes which may be linked to local deacetylation of histone H3. Represses HNF1A-mediated transcription. Required for the repression of ESRRG by CREBZF. Modulates AP-1 transcription factor activity. Deacetylates NR1H3 AND NR1H2 and deacetylation of NR1H3 at 'Lys-434' positively regulates transcription of NR1H3:RXR target genes, promotes NR1H3 proteosomal degradation and results in cholesterol efflux; a promoter clearing mechanism after reach round of transcription is proposed. Involved in lipid metabolism. Implicated in regulation of adipogenesis and fat mobilization in white adipocytes by repression of PPARG which probably involves association with NCOR1 and SMRT/NCOR2. Deacetylates ACSS2 leading to its activation, and HMGCS1. Involved in liver and muscle metabolism. Through deacteylation and activation of PPARGC1A is required to activate fatty acid oxidation in skeletel muscle under low-glucose conditions and is involved in glucose homeostasis. Involved in regulation of PPARA and fatty acid beta-oxidation in liver. Involved in positive regulation of insulin secretion in pancreatic beta cells in response to glucose; the function seems to imply transcriptional repression of UCP2. Proposed to deacetylate IRS2 thereby facilitating its insulin-induced tyrosine phosphorylation. Deacetylates SREBF1 isoform SREBP-1C thereby decreasing its stability and transactivation in lipogenic gene expression. Involved in DNA damage response by repressing genes which are involved in DNA repair, such as XPC and TP73, deacetylating XRCC6/Ku70, and faciliting recruitment of additional factors to sites of damaged DNA, such as SIRT1-deacetylated NBN can recruit ATM to initiate DNA repair and SIRT1-deacetylated XPA interacts with RPA2. Also involved in DNA repair of DNA double-strand breaks by homologous recombination and specifically single-strand annealing independently of XRCC6/Ku70 and NBN. Transcriptional suppression of XPC probably involves an E2F4:RBL2 suppressor complex and protein kinase B (AKT) signaling. Transcriptional suppression of TP73 probably involves E2F4 and PCAF. Deacetylates WRN thereby regulating its helicase and exonuclease activities and regulates WRN nuclear translocation in response to DNA damage. Deacetylates APEX1 at 'Lys-6' and 'Lys-7' and stimulates cellular AP endonuclease activity by promoting the association of APEX1 to XRCC1. Increases p53/TP53-mediated transcription-independent apoptosis by blocking nuclear translocation of cytoplasmic p53/TP53 and probably redirecting it to mitochondria. Deacetylates XRCC6/Ku70 at 'Lys-539' and 'Lys-542' causing it to sequester BAX away from mitochondria thereby inhibiting stress-induced apoptosis. Is involved in autophagy, presumably by deacetylating ATG5, ATG7 and MAP1LC3B/ATG8. Deacetylates AKT1 which leads to enhanced binding of AKT1 and PDK1 to PIP3 and promotes their activation. Proposed to play role in regulation of STK11/LBK1-dependent AMPK signaling pathways implicated in cellular senescence which seems to involve the regulation of the acetylation status of STK11/LBK1. Can deacetylate STK11/LBK1 and thereby increase its activity, cytoplasmic localization and association with STRAD; however, the relevance of such activity in normal cells is unclear. In endothelial cells is shown to inhibit STK11/LBK1 activity and to promote its degradation. Deacetylates SMAD7 at 'Lys-64' and 'Lys-70' thereby promoting its degradation. Deacetylates CIITA and augments its MHC class II transactivation and contributes to its stability. Deacteylates MECOM/EVI1. Deacetylates PML at 'Lys-487' and this deacetylation promotes PML control of PER2 nuclear localization. During the neurogenic transition, repress selective NOTCH1-target genes throug
    Isoform 2: Isoform 2 is shown to deacetylate 'Lys-382' of p53/TP53, however with lower activity than isoform 1. In combination, the two isoforms exert an additive effect. Isoform 2 regulates p53/TP53 expression and cellular stress response and is in turn repressed by p53/TP53 presenting a SIRT1 isoform-dependent auto-regulatory loop.
    (Microbial infection) In case of HIV-1 infection, interacts with and deacetylates the viral Tat protein. The viral Tat protein inhibits SIRT1 deacetylation activity toward RELA/NF-kappa-B p65, thereby potentiates its transcriptional activity and SIRT1 is proposed to contribute to T-cell hyperactivation during infection.
    SirtT1 75 kDa fragment: catalytically inactive 75SirT1 may be involved in regulation of apoptosis. May be involved in protecting chondrocytes from apoptotic death by associating with cytochrome C and interfering with apoptosome assembly.
  • 组织特异性Widely expressed.
  • 序列相似性Belongs to the sirtuin family. Class I subfamily.
    Contains 1 deacetylase sirtuin-type domain.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm. Mitochondrion and Nucleus, PML body. Cytoplasm. Nucleus. Recruited to the nuclear bodies via its interaction with PML (PubMed:12006491). Colocalized with APEX1 in the nucleus (PubMed:19934257). May be found in nucleolus, nuclear euchromatin, heterochromatin and inner membrane (PubMed:15469825). Shuttles between nucleus and cytoplasm (By similarity). Colocalizes in the nucleus with XBP1 isoform 2 (PubMed:20955178).
  • Information by UniProt
  • 数据库链接
  • 别名
    • 75SirT1 antibody
    • hSIR2 antibody
    • hSIRT1 antibody
    • HST2, S. cerevisiae, homolog of antibody
    • NAD dependent deacetylase sirtuin 1 antibody
    • NAD dependent protein 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
    see all

Anti-SIRT1 antibody [19A7AB4] 图像

  • Immunocytochemistry analysis using ab110304 at 0.5µg/ml staining SIRT1 in HDFn cells (paraformaldehyde fixed and Triton X-100 permeabilized). The secondary antibody was (Red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. 10% Goat serum was used as the blocking agent for all blocking steps.
  • IHC image of SIRT1 staining in Human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110304, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times

  • HL-60 cells were stained with 1 µg/mL SIRT1 antibody ab110304(blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.
  • All lanes : Anti-SIRT1 antibody [19A7AB4] (ab110304) at 1/1000 dilution

    Lane 1 : rat Skeletal muscle
    Lane 2 : rat Skeletal muscle

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat anti-Mouse at 1/6000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 81 kDa
    Additional bands at : 120 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 13 minutes

    This image is courtesy of an anonymous Abreview

    Blocked with 3% milk (TBS-tween) at 4C for 16 hours

    See Abreview

  • All lanes : Anti-SIRT1 antibody [19A7AB4] (ab110304) at 0.125 µg/ml

    Lane 1 : HepG2 cell lysate(Human)
    Lane 2 : H9C2 cell lysate (Rat)
    Lane 3 : MEF cell lysate (Mouse)


    Predicted band size : 81 kDa

    WB Conditions
    Primary Antibody: 0.25 µg/mL in 10X Blocking Buffer (ab126587). 3hrs at room temperature.

    Secondary Antibody: 1:5,000  in 10X Blocking Buffer (ab126587). 3hrs at room temperature.

    ab110304 detects a band of approximately 110 kDa (110-121 kDa) which is likely to be due to post translational glycosylation. SIRT1 is known to bind to several other proteins, and the 121kDa band could also be due to the presence of one of these complexes (ensure samples are adequately reduced and denatured).

Anti-SIRT1 antibody [19A7AB4] (ab110304)参考文献

This product has been referenced in:
  • Bellio MA  et al. Physiologic and Hypoxic Oxygen Concentration Differentially Regulate Human c-Kit+ Cardiac Stem Cell Proliferation and Migration. Am J Physiol Heart Circ Physiol N/A:ajpheart.00449.2016 (2016). WB ; Human . Read more (PubMed: 27694215) »
  • Xiao X  et al. GITR subverts Foxp3(+) Tregs to boost Th9 immunity through regulation of histone acetylation. Nat Commun 6:8266 (2015). WB ; Mouse . Read more (PubMed: 26365427) »

See all 12 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (AR42J-B13 Pancreatic tumour cell line)
Permeabilization Yes - 0.1% triton/PBS
Specification AR42J-B13 Pancreatic tumour cell line
Blocking step Roche Blocking reagent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Dr. Zoe Burke

Verified customer

提交于 May 12 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (8%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Mar 07 2016

Application Western blot
Sample Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions Reduced Denaturing (8%)
Loading amount 20 µg
Specification Skeletal muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Mar 07 2016

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Human Tissue lysate - whole (Skeletal Muscle)
Specification Skeletal Muscle
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Brittany Edgett

Verified customer

提交于 Jan 26 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (8% Gel)
Sample Mouse Tissue lysate - whole (Skeletal muscle (GTN))
Specification Skeletal muscle (GTN)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jun 02 2014

SIRT1 is widely expressed in majority of cell lines. HL-60 was selected for ease of preparation for flow cytometry (HL-60 is a suspension cell line; generally suspension cell lines easy to prepare for FC). For example:
http://www.proteinatlas.org/E...

Read More
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step Enzymatic
Sample Rat Tissue sections (Gonad)
Specification Gonad
Permeabilization No
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Oct 29 2013

Application Western blot
Loading amount 75 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Rat Cell lysate - other (Epithelial cells)
Specification Epithelial cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Oct 21 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (4-12)
Sample Rat Cell lysate - whole cell (Primary Neurons)
Specification Primary Neurons
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 Sep 05 2013

Thank you for contacting us.

I can confirm that ab6006 is suitable for use with antibodies ab119484, ab80039 and ab110304.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice...

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