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SIRT1 Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER130811

SIRT1 Rabbit Polyclonal Antibody

Application

  • WB

  • IHC-P

  • FC

  • IF-Cell

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

SIRT1 Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within Human SIRT1 aa 698-747 / 747.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, FC, IF-Cell

Molecular Weight

Predicted band size: 82 kDa

Positive Control

Hela cell lysate, Jurkat cell lysate, F9 cell lysate, HeLa, F9, human colon carcinoma tissue, human lung carcinoma tissue, mouse liver tissue, mouse testis tissue.

Conjugation

unconjugated

RRID

AB_3069022

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:200

  • FC

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

SirT1, the mammalian ortholog of Sir2, is a nuclear protein implicated in the regulation of many cellular processes, including apoptosis, cellular senescence, endocrine signaling, glucose homeostasis, aging, and longevity. Targets of SirT1 include acetylated p53, p300, Ku70, forkhead (FoxO) transcription factors, PPARγ, and the PPARγ coactivator-1α (PGC-1α) protein. Deacetylation of p53 and FoxO transcription factors represses apoptosis and increases cell survival. SirT1 deacetylase activity is inhibited by nicotinamide and activated by resveratrol.

Background References

1. Yang Y et al. Regulation of SIRT1 and Its Roles in Inflammation. Front Immunol. 2022 Mar

2. Lu C et al. Novel Role of the SIRT1 in Endocrine and Metabolic Diseases. Int J Biol Sci. 2023 Jan

Sequence Similarity

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

Nucleus, cytoplasm, Mitochondrion.

UNIPROT

SwissProt: Q96EB6 Human

Synonyms

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

Expand

Images

  • Western blot analysis of SIRT1 on different lysates with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/500 dilution.<br /><br />Lane 1: Hela cell lysate<br />Lane 2: Jurkat cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 82 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 2 minutes;<br /><br />6% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SIRT1 on different lysates with Rabbit anti-SIRT1 antibody (ER130811) at 1/500 dilution.

    Lane 1: Hela cell lysate
    Lane 2: Jurkat cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 82 kDa
    Observed band size: 110 kDa

    Exposure time: 2 minutes;

    6% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER130811) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of SIRT1 on F9 cell lysates with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/2,000 dilution.<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 82 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 1 minute;<br /><br />8% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SIRT1 on F9 cell lysates with Rabbit anti-SIRT1 antibody (ER130811) at 1/2,000 dilution.

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 82 kDa
    Observed band size: 110 kDa

    Exposure time: 1 minute;

    8% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER130811) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling SIRT1 with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling SIRT1 with Rabbit anti-SIRT1 antibody (ER130811) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SIRT1 antibody (ER130811) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of F9 cells labeling SIRT1 with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of F9 cells labeling SIRT1 with Rabbit anti-SIRT1 antibody (ER130811) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SIRT1 antibody (ER130811) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-SIRT1 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-SIRT1 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-SIRT1 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-SIRT1 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-SIRT1 antibody (ER130811) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER130811) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-SIRT1 antibody (ER130811) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER130811) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HeLa cells labeling SIRT1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling SIRT1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ER130811, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Flow cytometric analysis of F9 cells labeling SIRT1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of F9 cells labeling SIRT1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ER130811, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of SIRT1 on different lysates with Rabbit anti-SIRT1 antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/20,000 dilution.<br /><br />Lane 1: HEK-293-si NT cell lysate<br />Lane 2: HEK-293-si SIRT1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 82 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 50 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER130811" style="font-weight: bold;text-decoration: underline;">ER130811</a>) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/100,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of SIRT1 on different lysates with Rabbit anti-SIRT1 antibody (ER130811) at 1/20,000 dilution.

    Lane 1: HEK-293-si NT cell lysate
    Lane 2: HEK-293-si SIRT1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 82 kDa
    Observed band size: 110 kDa

    Exposure time: 50 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER130811) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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