1. Antibodies
  2. Homo sapiens
  3. RUNX3

ENCAB000AKM

Antibody against Homo sapiens RUNX3

Homo sapiens
at least one cell type or tissue
awaiting characterization
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-101553
Lot ID
B0909
Characterized targets
RUNX3 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Isotype
IgG
Antigen description
Raised against amino acids 191-300 of recombinant RUNX3 of human origin.

Characterizations

RUNX3 (Homo sapiens)
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
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 InvitrogenBenchPro 4100 system. Visualization was achieved using SuperSignal West Femto solution (Thermo Scientific).Results: Band of expected size visualized, representing strongest signal in the lane.The ~50 kDa and ~45 kDa bands were both tested with IP-mass spec.Only the ~45kDa band was positive for RUNX3.See validation 2. Figure legend: IP-western with sc-101553 in whole cell lysate (WCL) of GM12878; PM=protein marker. RUNX3 band is indicated.
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB
RUNX3 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment from submitter
Attachment from submitters
not reviewed
Caption
IP followed by mass spectrometry: Briefly, GM12878 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 ofAlabama 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_RUNX3_ss101553_09262011_MassSpec.pdf), including common contaminants. Target protein, RUNX3, is listed as hit 3a in the ~45 kDa band.The ~50 kDa band shown in Validation 1 was also tested by mass spec and was positive for heavy chain, but not for RUNX3.
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB