1. Antibodies
  2. Homo sapiens
  3. MBD4

ENCAB000AIN

Antibody against Homo sapiens MBD4

Homo sapiens
at least one cell type or tissue
awaiting characterization
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-271530
Lot ID
A1111
Characterized targets
MBD4 (Homo sapiens)
Lot ID aliases
H1210
Host
mouse
Clonality
monoclonal
Isotype
IgG
Antigen description
Epitope mapping between a.a. 515-555 within an internal region of MBD4 of mouse origin.

Characterizations

MBD4 (Homo sapiens)
Method: immunoblot
Attachment PDF Icon
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. Lower band also visualized at ~58 kDa. Both bands tested by IP-mass spec, but only the ~75 kDa band was positive for MBD4 (see below). Figure legend: IP-western with sc-271530 in whole cell lysate (WCL) of K562 and GM12878; PM=protein marker. MBD4 band is indicated.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
MBD4 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
not reviewed
Caption
IP followed by mass spectrometry: Briefly, K562 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. 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_MBD4_sc271530_08252011_MassSpec.pdf), including common contaminants. Target protein is listed as hit 9 in the upper (~75kDa) band and absent from the lower (~58kDa) band.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB