ENCAB000AJE

Antibody against Homo sapiens NFATC1

Homo sapiens
at least one cell type or tissue
awaiting characterization
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-17834
Lot ID
G2007
Characterized targets
NFATC1 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Isotype
IgG
Antigen description
Raised against amino acids 1-110 of NFATc1 of human origin. Antibody Target: NFATC1

Characterizations

NFATC1 (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: Bands of expected size visualized, representing strongest signal in the lane. Figure legend: IP-western with sc-17834 in nuclear extract (NE) of K562 and whole cell lysate (WCL) of K562, GM12878 and HepG2; PM=protein marker. NFATC1 bands are indicated.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
NFATC1 (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 (lower bands at ~85-100kDa, and upper bands at ~105-200kDa) 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_NFATC1_sc17834_09012011_MassSpec.pdf), including common contaminants. Target protein is listed as hit 16a in lower (~85-100 kDa) bands and 27a in upper (~105-200 kDa) bands.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB