ENCODE DCC Antibody Validation Document

Transcription

1 ENCODE DCC Antibody Validation Document Date of Submission Name: Lab Antibody Name: Target: Company/ Source: Catalog Number, database ID, laboratory Lot Number Antibody Description: Target Description: Species Target Species Host Validation Method #1 Validation Method #2 Purification Method Polyclonal/ Monoclonal Vendor URL: Reference (PI/ Publication Information) Please complete the following for antibodies to histone modifications: if your specifications are not listed in the drop-down box, please write-in the appropriate information Histone Name AA modified AA Position Modification

2 Validation #1 Analysis Insert Validation Image (click here)

3 Valida&on 1: Immunoblot/immunoprecipita&on with sc 372 (NFKB p65 ) Immunoprecipita-ons using sc 372 were carried out in GM12878 cells treated with TNFa (6 hrs), murine CH12 cells and murine MEL cells. NE= nuclear lysate, S= supernatant from immunoprecipita-on with sc 372, IP= bound material from immunoprecipita-on with sc 372, IgG= bound material from immunoprecipita-on with control IgG. Arrows indicate band consistent with the expected size of NFKB p65. Bands indicated by * in GM12878 immunoprecipita-on are IgG heavy and light chains. These bands are absent from CH12 and MEL immunoprecipita-ons because an-body was crosslinked to Protein A beads prior to use in immunoprecipita-on. Molecular weights are indicated in kd. GM12878 (+TNFα) NE S IP IgG CH12 MEL 1 75 * 25 * *

4 Validation #2 Analysis Insert Validation Image (Click here)

5 Table 1. NFKB mo#f enrichment. Cell Line Motif Enrichment (log2) Enrichment p-value (-log10) Fraction of peaks containing motif NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known5) (NF-kappaB_known5) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) NFKB_GM (NF-kappaB_known1) (NF-kappaB_known1) (NF-kappaB_known5) Figure 2. Mo#f consensus sequences and posi#on weight matrices for highly enriched NFKB mo#f. NFKB_known1 NFKB_known5 Posi9on/ Consensus >NF-kappaB_known1 NF-kappaB-(p)_transfac_M A C G T G A K Y C C Posi9on/ Consensus >NF-kappaB_known5 RELA_jaspar_MA A C G T B G R V T T T