ENCAB000AMB

Antibody against Homo sapiens TRIM28

Homo sapiens
at least one cell type or tissue
awaiting characterization
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-81411
Lot ID
L0309
Characterized targets
TRIM28 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Isotype
IgG
Antigen description
Raised against a recombinant protein corresponding to an internal region of TRIM28 of human origin.

Characterizations

TRIM28 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
not reviewed
Caption
IP followed by mass spectrometry: 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 Coomassie Blue in order to visualize marker bands. Gel fragments corresponding to the bands indicated above in the western blot image were excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. There the samples were run on an LTQ XL Linear Ion Trap Mass Spectrometer by LC-ESI-MS/MS. Peptides were identified using SEQUEST tandem mass spectra analysis, with probability based matching at p < 0.05. As per ENCODE data standards, all SEQUEST results are attached (ENCODE_HAIB_TRIM28_sc81411_20012012_MassSpec.pdf), including common contaminants. Target protein is listed as hit 1 in the ~100 kDa band.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
TRIM28 (Homo sapiens)
Method: immunoblot
not reviewed
Caption
Western blot protocol: Whole cell lysate was 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. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen iBlot system. Blotting with primary (same as that used for IP) and secondary HRP-conjugated antibodies was performed on an Invitrogen BenchPro 4100 system. Visualization was achieved using SuperSignal West Femto solution (Thermo Scientific). Results: Band of expected size (~100 kDa) visualized, and tested positive for TRIM28 by IP-mass spec (see Validation 2). Figure legend: IP-western with sc-81411 in whole cell lysate (WCL) of K562, GM12878, HepG2 and HeLa; PM=protein marker. TRIM28 bands are indicated.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB