1. Antibodies
  2. Homo sapiens
  3. CREB1

ENCAB000AFN

Antibody against Homo sapiens CREB1

Homo sapiens
at least one cell type or tissue
awaiting characterization
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-240
Lot ID
C2306
Characterized targets
CREB1 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Isotype
IgG1
Antigen description
Raised against amino acids 254-327 mapping within the DNA binding and dimerization domain of human CREB-1.

Characterizations

CREB1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
not reviewed
Caption
IP followed by mass spectrometry: Briefly, A549 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_CREB1_sc240_09282011_MassSpec.pdf), including common contaminants. Target protein is listed as hit 8a in the ~50kDa band.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
CREB1 (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. Figure legend: IP-western with sc-240 in whole cell lysate (WCL) of A549; PM=protein marker. CREB1 band is indicated.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB