SIRT6 antibody | knockout validation | Cell Signaling Technology 12486

This is a knockout-validated antibody summary, based on the publication "SIRT6 safeguards human mesenchymal stem cells from oxidative stress by coactivating NRF2" [1] (as discussed below) and article "SIRT6 regulates Ras-related protein R-Ras2 by lysine defatty-acylation" [2] (see figure 4a in the article). Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

SIRT6 antibody | knockout validation | Cell Signaling Technology 12486 figure 1
Figure 1. Western blotting analysis of SIRT6 protein in WT and SIRT6-deficient hMSCs. β-actin was used as the loading control. From [1].
Antibody information

Rabbit monoclonal

Company: Cell Signaling Technology

Antibody: SIRT6

Catalog number: 12486

Summary: Rabbit monoclonal against full-length mouse SirT6 protein. Reacts with human, mouse, rat,and monkey, and is suitable for western blotting, immunoprecipitation and immunofluorescence (immunocytochemistry).

Validation Method

Western blot | Immunocytochemistry

SIRT6 antibody | knockout validation | Cell Signaling Technology 12486 figure 2
Figure 2. Immunofluorescence analysis showing the absence of SIRT6 protein in the nuclei of SIRT6-deficient hMSCs. Scale bar, 10 μm. From [1].
Sample

WB and IC: wild-type and SIRT6-deficient human embryonic stem (hESC) cells and human mesenchymal stem cells (hMSC).

IC: hESCs and hMSCs were fixed with formaldehyde (4% in PBS) for 15 min, permeabilized with Triton X-100 (0.4% in PBS) for 15 min.

Blocking agent

IC: 10% donkey serum in PBS for 30 min.

Primary incubation

WB: 1:1,000

IC: 1:100, overnight at 4 °C.

Secondary incubation

IC: for 1 h at room temperature.

References
  1. Pan H, Guan D, Liu X, Li J, Wang L, Wu J, et al. SIRT6 safeguards human mesenchymal stem cells from oxidative stress by coactivating NRF2. Cell Res. 2016;26:190-205 pubmed publisher
  2. Zhang X, Spiegelman N, Nelson O, Jing H, Lin H. SIRT6 regulates Ras-related protein R-Ras2 by lysine defatty-acylation.. elife. 2017;6: pubmed publisher