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SIRT3 Recombinant Rabbit Monoclonal Antibody [PSH05-22]

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Specification

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Catalog# HA722251

SIRT3 Recombinant Rabbit Monoclonal Antibody [PSH05-22]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

  • IF-Tissue

  • IP

  • ChIP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

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

Overview

Product Name

SIRT3 Recombinant Rabbit Monoclonal Antibody [PSH05-22]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human SIRT3 aa 51-399 / 399.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC, IF-Tissue, IP, ChIP

Molecular Weight

Predicted band size: 44 kDa

Positive Control

PANC-1 cell lysate, HepG2 cell lysate, A549 cell lysate, SiHa cell lysate, mouse liver tissue lysate, mouse kidney tissue lysate, rat liver tissue lysate, human liver tissue, human colon tissue, HeLa cell lysate, HEK-293 cell lysate, SW480 cell lysate, HT-29 cell lysate, rat kidney tissue lysate, human kidney tissue, mouse liver tissue, rat liver tissue, HepG2, NIH/3T3.

Conjugation

unconjugated

Clone Number

PSH05-22

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

  • IF-Tissue

  • 1:200

  • IP

  • 1:2000

  • ChIP

  • Use 0.5~2 μg for 25 μg of chromatin.

Target

Function

NAD-dependent deacetylase sirtuin-3, mitochondrial also known as SIRT3 is a protein that in humans is encoded by the SIRT3 gene. SIRT3 is member of the mammalian sirtuin family of proteins, which are homologs to the yeast Sir2 protein. SIRT3 exhibits NAD+-dependent deacetylase activity. Members of the sirtuin family are characterized by a sirtuin core domain and grouped into four classes, and the protein encoded by this gene is included in class I of the sirtuin family. The human sirtuins have a range of molecular functions and have emerged as important proteins in aging, stress resistance, and metabolic regulation. Yeast sirtuin proteins are known to regulate epigenetic gene silencing and suppress recombination of rDNA. In addition to protein deacetylation, studies have shown that the human sirtuins may also function as intracellular regulatory proteins with mono ADP ribosyltransferase activity.

Background References

1. Zhou L et al. The Role of SIRT3 in Exercise and Aging. Cells. 2022 Aug

2. Diao Z et al. SIRT3 consolidates heterochromatin and counteracts senescence. Nucleic Acids Res. 2021 May

Subcellular Location

Mitochondrion matrix.

UNIPROT

SwissProt: Q9NTG7 Human

SwissProt: Q8R104 Mouse

Entrez Gene: 293615 Rat

Synonyms

hSIRT 3 antibody

hSIRT3 antibody

Mitochondrial nicotinamide adenine dinucleotide dependent deacetylase antibody

NAD dependent deacetylase sirtuin 3 mitochondrial antibody

NAD-dependent protein deacetylase sirtuin-3, mitochondrial antibody

Regulatory protein SIR2 homolog 3 antibody

Silent mating type information regulation 2 S.cerevisiae homolog 3 antibody

Sir 2 like 3 antibody

SIR 2 like protein 3 antibody

SIR 3 antibody

Expand

Images

  • Western blot analysis of SIRT3 on different lysates with Rabbit anti-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.<br /><br />Lane 1: PANC-1 cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: A549 cell lysate<br />Lane 4: SiHa cell lysate<br />Lane 5: Mouse liver tissue lysate<br />Lane 6: Mouse kidney tissue lysate<br />Lane 7: Rat liver tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 44 kDa<br />Observed band size: 28 kDa<br /><br />Exposure time: 30 seconds; ECL: K1801;<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/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SIRT3 on different lysates with Rabbit anti-SIRT3 antibody (HA722251) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

    Lane 1: PANC-1 cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: A549 cell lysate
    Lane 4: SiHa cell lysate
    Lane 5: Mouse liver tissue lysate
    Lane 6: Mouse kidney tissue lysate
    Lane 7: Rat liver tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 44 kDa
    Observed band size: 28 kDa

    Exposure time: 30 seconds; ECL: K1801;

    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 (HA722251) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 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 human liver tissue with Rabbit anti-SIRT3 antibody (HA722251) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722251) at 1/1,000 dilution and competitor's antibody at 1/1,000 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 human colon tissue with Rabbit anti-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 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 human colon tissue with Rabbit anti-SIRT3 antibody (HA722251) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722251) at 1/1,000 dilution and competitor's antibody at 1/1,000 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.

  • Western blot analysis of SIRT3 on different lysates with Rabbit anti-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HEK-293 cell lysate (20 µg/Lane)<br />Lane 3: PANC-1 cell lysate (20 µg/Lane)<br />Lane 4: HepG2 cell lysate (20 µg/Lane)<br />Lane 5: SW480 cell lysate (20 µg/Lane)<br />Lane 6: A549 cell lysate (20 µg/Lane)<br />Lane 7: HT-29 cell lysate (20 µg/Lane)<br />Lane 8: SiHa cell lysate (20 µg/Lane)<br />Lane 9: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 10: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 11: Rat liver tissue lysate (40 µg/Lane)<br />Lane 12: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 44 kDa<br />Observed band size: 28 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<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/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/2,000 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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of SIRT3 on different lysates with Rabbit anti-SIRT3 antibody (HA722251) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HEK-293 cell lysate (20 µg/Lane)
    Lane 3: PANC-1 cell lysate (20 µg/Lane)
    Lane 4: HepG2 cell lysate (20 µg/Lane)
    Lane 5: SW480 cell lysate (20 µg/Lane)
    Lane 6: A549 cell lysate (20 µg/Lane)
    Lane 7: HT-29 cell lysate (20 µg/Lane)
    Lane 8: SiHa cell lysate (20 µg/Lane)
    Lane 9: Mouse liver tissue lysate (40 µg/Lane)
    Lane 10: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 11: Rat liver tissue lysate (40 µg/Lane)
    Lane 12: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 44 kDa
    Observed band size: 28 kDa

    Exposure time: 25 seconds; ECL: K1801;

    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 (HA722251) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of SIRT3 on different lysates with Rabbit anti-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-SIRT3 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 44 kDa<br />Observed band size: 28 kDa<br /><br />Exposure time: 2 minutes; ECL: K1801;<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/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of SIRT3 on different lysates with Rabbit anti-SIRT3 antibody (HA722251) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-SIRT3 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 44 kDa
    Observed band size: 28 kDa

    Exposure time: 2 minutes; ECL: K1801;

    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 (HA722251) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 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 human kidney tissue with Rabbit anti-SIRT3 antibody (HA722251) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722251) at 1/1,000 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-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 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-SIRT3 antibody (HA722251) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722251) at 1/1,000 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 rat liver tissue with Rabbit anti-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) at 1/1,000 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 rat liver tissue with Rabbit anti-SIRT3 antibody (HA722251) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722251) at 1/1,000 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.

  • Immunocytochemistry analysis of HepG2 cells labeling SIRT3 with Rabbit anti-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</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-SIRT3 antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</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. Counterstained with Mitotracker (Red). Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HepG2 cells labeling SIRT3 with Rabbit anti-SIRT3 antibody (HA722251) 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-SIRT3 antibody (HA722251) 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. Counterstained with Mitotracker (Red). Nuclear DNA was labelled in blue with DAPI.

  • Flow cytometric analysis of HepG2 cells labeling SIRT3.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>, 1μg/mL) (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 HepG2 cells labeling SIRT3.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722251, 1μg/mL) (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 NIH/3T3 cells labeling SIRT3.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>, 1μg/mL) (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 NIH/3T3 cells labeling SIRT3.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722251, 1μg/mL) (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).

  • SIRT3 was immunoprecipitated from 0.2 mg HepG2 cell lysate with <a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a> at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HepG2 cell lysate (input)<br />Lane 2: <a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a> IP in HepG2 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a> in HepG2 cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute 17 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    SIRT3 was immunoprecipitated from 0.2 mg HepG2 cell lysate with HA722251 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722251 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HepG2 cell lysate (input)
    Lane 2: HA722251 IP in HepG2 cell lysate
    Lane 3: Rabbit IgG instead of HA722251 in HepG2 cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute 17 seconds; ECL: K1801

  • Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with SIRT3 (<a href="/products/HA722251" style="font-weight: bold;text-decoration: underline;">HA722251</a>) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with SIRT3 (HA722251) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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