1. Antibodies
  2. Homo sapiens
  3. CTCF

ENCAB000AXX

Antibody against Homo sapiens CTCF

Homo sapiens
HEK293, A549, OCI-LY1, MM.1S, MCF-7, K562, DOHH2, bipolar neuron, A673, SK-N-SH, smooth muscle cell, Loucy, fibroblast of arm, myotube, OCI-LY3, cardiac muscle cell, hepatocyte, SU-DHL-6, neutrophil, GM23338, PC-9, PC-3, astrocyte, neural progenitor cell, fibroblast of dermis, KMS-11, NCI-H929, HCT116, OCI-LY7, spleen, stomach, tibial nerve, upper lobe of left lung, uterus, adrenal gland, ascending aorta, gastroesophageal sphincter, Peyer's patch, esophagus muscularis mucosa, prostate gland, right atrium auricular region, gastrocnemius medialis, thyroid gland, GM23248, Karpas-422, coronary artery, vagina, heart left ventricle, testis, parathyroid adenoma, body of pancreas, esophagus squamous epithelium, transverse colon
characterized to standards with exemption
Status
released
Source (vendor)
Millipore
Product ID
07-729
Lot ID
DAM1472197
Characterized targets
CTCF (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
crude
Antigen description
Raised against KLH-conjugated, synthetic peptide corresponding to amino acids 659-675 ( C-TNQPKQNQPTAIIQVED) of human CCCTC-binding factor (CTCF) with a N-terminal cysteine added for cunjugation purposes
Antigen sequence
C-TNQPKQNQPTAIIQVED
Aliases
bradley-bernstein:PchAb 51-V

Characterizations

CTCF (Homo sapiens)
HEK293
Method: immunoblot
Attachment from submitter
Attachment from submitters
compliant
Caption
5X10e5 cells equivalent of nuclear extract from HEK293 cells, were resolved by electrophoresis on a 4-12% acrylamide gel. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen iBlot system. Membrane was blocked for an hour in room temperature, with 5% nonfat dry milk and blotted with primary antibody in the appropriate concentration over night at 4c. Membrane was washed and blotted with secondary HRP-conjugated antibody. Detection was made with Optiblot ECL Detect Kit (ab133406) for 2 min. Band of the expected size was detected (~120kDa), the primary reactive band contains at least 50% of the signal observed on the blot.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
CTCF (Homo sapiens)
Method: ChIP-seq comparison
Attachment PDF Icon
exempt from standards
Caption
This validation relies on the use of two different antibodies raised against different, non-overlapping peptides, both derived from the sequence of human CTCF, and the discovery of highly correlated patterns of enrichment by ChIP-Seq in K562 cells. The polyclonal antibody being validated (ENCAB000AXX) was raised against a 17 residue long synthetic peptide located 52 residues away from the C-terminus of CTCF. In contrast, the monoclonal antibody (https://www.encodeproject.org/antibodies/ENCAB498PCM/) was raised against a peptide derived from the extreme C-terminus of CTCF. Overall correlation score = 0.976
Submitter comment
SAV5
Reviewer comment
There is no more antibody to characterize. This method was acceptable for ENCODE2.
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad
CTCF (Homo sapiens)
A549OCI-LY1MM.1SMCF-7K562DOHH2bipolar neuronA673SK-N-SHsmooth muscle cellLoucyfibroblast of armmyotubeOCI-LY3cardiac muscle cellhepatocyteSU-DHL-6neutrophilGM23338PC-9PC-3astrocyteneural progenitor cellfibroblast of dermisKMS-11NCI-H929HCT116OCI-LY7spleenstomachtibial nerveupper lobe of left lunguterusadrenal glandascending aortagastroesophageal sphincterPeyer's patchesophagus muscularis mucosaprostate glandright atrium auricular regiongastrocnemius medialisthyroid glandGM23248Karpas-422coronary arteryvaginaheart left ventricletestis
Method: immunoblot
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Both ENCODE submitter and reviewer have agreed that this is a well-established antibody lot. Hence it does not require additional validation as standardized by the ENCODE Consortium.
Submitter comment
The lab requests that this is considered a well-documented antibody lot and therefore does not require additional validation.
Reviewer comment
Dr. Peggy J Farnham: I would support a “no additional validation needed” classification for this antibody.
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad
Download
 exempted.png
CTCF (Homo sapiens)
parathyroid adenomabody of pancreasesophagus squamous epitheliumtransverse colon
Method: immunoblot
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
The ENCODE Binding Working Group finds for some valuable tissues that recreating a primary on well characterized antibodies is not cost effective. Therefore, they allow exemption from standards for these tissues. Compliant characterizations in other cell lines may be used instead.
Submitter comment
--
Reviewer comment
--
Submitted by
Kathrina Onate
Lab
Bradley Bernstein, Broad
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 No_tissue.png