1. Antibodies
  2. Homo sapiens
  3. ZEB1

ENCAB000AMO

Antibody against Homo sapiens ZEB1

Homo sapiens
GM12878, bipolar spindle neuron, bipolar neuron
characterized to standards with exemption
Homo sapiens
HepG2
not characterized to standards
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-25388
Lot ID
D2010
Characterized targets
ZEB1 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Isotype
IgG
Antigen description
Epitope corresponding to amino acids 39-140 mapping near the N-terminus of ZEB1 of human origin.

Characterizations

ZEB1 (Homo sapiens)
GM12878
Method: immunoblot
Attachment PDF Icon
exempt from standards
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-25388 in whole cell lysate (WCL) of GM12878. ZEB1 bands are indicated, as is heavy chain of IgG.
Submitter comment
This is an ENCODE2 antibody that there is no more of to bring up to date.
Reviewer comment
This would have met the ENCODE2 standards.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
ZEB1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
exempt from standards
Caption
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 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, including common contaminants. Target protein is listed as hit 16 for lower band and hit 14 for upper band (both hits highlighted in bold font).
Submitter comment
This is an ENCODE2 antibody that there is no more of to bring up to date.
Reviewer comment
This would have passed ENCODE2 standards
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
ZEB1 (Homo sapiens)
bipolar spindle neuronbipolar neuron
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
The ENCODE Binding Working Group finds for some valuable biosamples that recreating a primary on well characterized antibodies is not cost effective. Therefore, they allow exemption from standards for these samples.
Submitter comment
The lab is asking for an exemption for IPSCs and biopolar spindle neurons (IPSC-derived) cells due to the lack of resources to make a primary characterization for them
Reviewer comment
Exempted by the Feb 29, 2016 antibody review panel, pending valid secondary charaacterization
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
ZEB1 (Homo sapiens)
GM12878HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
Whole cell lysates of GM12878 and HepG2 were immunoprecipitated using the primary antibody (Santa Cruz Biotechnology; sc-25388). The IP fraction was separated on a 12% acrylamide gel with the Bio-Rad PROTEAN II xi system. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen iBlot system. The membrane was probed with the primary antibody (same as that used for IP) and a secondary HRP-conjugated antibody. The resulting bands were visualized with SuperSignal West Femto Solution (Thermo Scientific). Protein Marker (PM) is labeled in kDa. The approximate size of ZEB1 is ~124 kDa.
Reviewer comment
Band of interest is not 50% of overall signal and is not within 20% of expected size
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB