ENCAB000AHJ

Homo sapiens

at least one cell type or tissue

awaiting characterization

Status: released
Source (vendor): Santa Cruz Biotech
Product ID: sc-6296
Lot ID: G0307
Characterized targets: HDAC2 (Homo sapiens)
Host: goat
Clonality: polyclonal
Isotype: IgG
Antigen description: Epitope mapping at the C-terminus of HDAC2 of human origin.
External resources: AR:AB_2248376
UCSC-ENCODE-cv:HDAC2_(SC-6296)

Method: immunoprecipitation followed by mass spectrometry

not reviewed

Caption: ENCODE data standards recognizes various methodologies for secondary validation of antibodies. Among these methodologies is immunoprecipitation followed by mass spectrometry analysis. Briefly, K562 whole cell lysates were immunoprecipitated using primary antibody, and the IP fraction was loaded on a 12% acrylamide gel and separated with a Bio-Rad PROTEAN II xi system. Gel was stained with Coomasie Blue in order to visualize marker bands. A gel fragment corresponding to the band indicated above in the western blot image was excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. There the sample was run on an LTQ XL Linear Ion Trap Mass Spectrometer with alternating collision-induced dissociation and electron-transfer dissociation. Peptides were identified using MASCOT (Matrix Science), with probability based matching at p < 0.05. Subsequent analysis was performed in Scaffold (Proteome Software, Inc.) at 0.0% protein FDR and 0.0% peptide FDR. As per ENCODE data standards, all Scaffold results are listed below, including common contaminants. Target protein is highlighted in bold font.
Submitted by: Richard Myers
Lab: Richard Myers, HAIB
Grant: U54HG004576
Download: human_HDAC2_SC-6296_validation_Myers.pdf

Method: immunoprecipitation

not reviewed