ENCAB273TMQ

Homo sapiens

any cell type or tissue

characterized to standards

Method: dot blot assay

compliant

Caption: We performed a DB analysis using the large set of modified peptides provided in the Active Motif MODified Histone Peptide Array, catalog #13005. The only peptide recognized by the H3K27me3 antibody was the correct one.
Submitted by: Nina Farrell
Lab: Bradley Bernstein, Broad
Grant: UM1HG009390
Download: H3K27me3_Thermo Fisher_MA533081_VB2948733_DB_ PchAb 1426.pdf

K562

Method: immunoblot

compliant

Caption: We performed a WB using K562 nuclear extract from K562 (15ug) cells . Samples were resolved by electrophoresis on a 4-12% Bis-Tris gel in 1X MES running buffer. After separation, the samples were transferred to a nitrocellulose membrane using an Invitrogen iBlot system (P3 7')' Membrane was blocked in TBST w/ 5% NFDM for 1 hr @ RT. Next, membrane was blotted with primary antibody at the appropriate concentration in blocking solution O/N @ 4°C. Membrane was washed and blotted with secondary HRP-conjugated antibody appropriately diluted in blocking buffer. Detection was made using an ECL Detection Kit and incubated for 0.5 min. Bands of expected size (~17kDa) were detected using a ChemiDoc camera and visualized in extract. Antibody seems to be specific since no reactivity with non histone proteins or unmodified histones was detected.
Submitted by: Nina Farrell
Lab: Bradley Bernstein, Broad
Grant: UM1HG009390
Download: H3K27me3_Thermo Fisher_MA533081_VB2948733_WB_ PchAb 1426.pdf