ENCAB000AIG

Antibody against Homo sapiens LEF1

Homo sapiens
at least one cell type or tissue
awaiting characterization
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-8592
Lot ID
C2409
Characterized targets
LEF1 (Homo sapiens)
Host
goat
Clonality
polyclonal
Isotype
IgG
Antigen description
Epitope mapping near the C-terminus of LEF1 of human origin.

Characterizations

LEF1 (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_LEF1_sc8592 _09122011_MassSpec.pdf), including common contaminants. Target protein is listed as hit 8 in the ~54 kDa band.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
LEF1 (Homo sapiens)
Method: immunoprecipitation
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 visualized, representing strongest signal in the lane. Figure legend: IP-western with sc-8592 in nuclear extract (NE) and whole cell lysate (WCL) of K562 and GM12878; PM=protein marker. LEF1 bands are indicated.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB