For western blots, cells were lysed in RIPA buffer (50 mM Tris-HCl, pH 7.4, 250 mM NaCl, 2% Nonidet-P40, 2.5 mM EDTA, 0.1% SDS,0.5% DOC) supplemented with PIC and PMSF, resolved on 6%–10% SDS-PAGE (Mini-PROTEAN electrophoresis chamber, Bio-Rad), and transferred on PVDF membranes (Mini Trans-Blot apparatus, Bio-Rad) following manufacturer’s protocols. Membranes were blocked in 5% milk in PBS with 0.1% tween (PBST) and incubated overnight at 4°C, or for 1hr at room temperature with primary antibodies (Rinf 84546S, CST 1:1000; Nanog A300–397A, Bethyl Laboratories 1:2000; Oct4 SC-5279, SantaCruz 1:500; H3 ab1791, abcam 1:15000; Tet2 ab124297, abcam 1:1000; Flag 14739S, CST 1:1000; Actin AC-15, abcam 1:40000). Secondary antibody incubations (HRP-anti mouse, 401253, or anti rabbit, 401393, 1:5000, CalBiochem) were carried out for 1hr at room temperature. For quantifying Rinf protein levels in chromatin bound and soluble fractions of the cell, ESC lysate was fractionated as described before (Zhang et al., 2016 (link)). Each fraction was analyzed by western blot using anti-Rinf antibody. In all experiments Actin or H3 were used as loading controls.
Tet2 ab124297
Manufactured by Abcam
Tet2 ab124297 is a recombinant rabbit monoclonal antibody that recognizes the Tet2 protein. Tet2 is a member of the ten-eleven translocation (TET) family of proteins, which are involved in DNA demethylation. This antibody can be used in various applications to detect and study the Tet2 protein.
Automatically generated - may contain errors
Lab products found in correlation
Oct4 sc 5279, by Santa Cruz Biotechnology (2 mentions)
Nanog a300 397a, by Fortis Life Sciences (2 mentions)
Rinf 84546s, by Fortis Life Sciences (2 mentions)
Flag 14739s, by Abcam (2 mentions)
Mini protean electrophoresis chamber, by Bio-Rad (2 mentions)
Mini trans blot apparatus, by Bio-Rad (2 mentions)
Actin ac 15, by Abcam (2 mentions)
H3 ab1791, by Abcam (2 mentions)
2 protocols using tet2 ab124297
1
Western Blot Analysis of Pluripotency Markers
2
Western Blot Analysis of Pluripotency Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For western blots, cells were lysed in RIPA buffer (50 mM Tris-HCl, pH 7.4, 250 mM NaCl, 2% Nonidet-P40, 2.5 mM EDTA, 0.1% SDS,0.5% DOC) supplemented with PIC and PMSF, resolved on 6%–10% SDS-PAGE (Mini-PROTEAN electrophoresis chamber, Bio-Rad), and transferred on PVDF membranes (Mini Trans-Blot apparatus, Bio-Rad) following manufacturer’s protocols. Membranes were blocked in 5% milk in PBS with 0.1% tween (PBST) and incubated overnight at 4°C, or for 1hr at room temperature with primary antibodies (Rinf 84546S, CST 1:1000; Nanog A300–397A, Bethyl Laboratories 1:2000; Oct4 SC-5279, SantaCruz 1:500; H3 ab1791, abcam 1:15000; Tet2 ab124297, abcam 1:1000; Flag 14739S, CST 1:1000; Actin AC-15, abcam 1:40000). Secondary antibody incubations (HRP-anti mouse, 401253, or anti rabbit, 401393, 1:5000, CalBiochem) were carried out for 1hr at room temperature. For quantifying Rinf protein levels in chromatin bound and soluble fractions of the cell, ESC lysate was fractionated as described before (Zhang et al., 2016 (link)). Each fraction was analyzed by western blot using anti-Rinf antibody. In all experiments Actin or H3 were used as loading controls.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!