ENCAB000AMD

Antibody against Homo sapiens UBTF, Mus musculus UBTF

Homo sapiens
K562, HeLa-S3, HepG2
characterized to standards
Homo sapiens
any cell type or tissue
partially characterized
Mus musculus
any cell type or tissue
partially characterized
Homo sapiens
GM12878
not characterized to standards
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-13125
Lot ID
B1804
Host
mouse
Clonality
monoclonal
Purification
other
Antigen description
UBF (F-9) is raised against amino acids 1-220 of UBF of human origin.

Characterizations

UBTF (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody sc-13125. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 97.0
Reviewer comment
Band appears in the IgG control.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
UBTF (Homo sapiens)
Method: ChIP-seq comparison
not reviewed
Caption
Using antibody SAB1404509 or sc-13125, we observe a major band in Western blots on lysates from cell lines K562, HeLa S3, HepG2, and, at lower intensity, GM12878 that migrates at a position consistent with the predicted size (~89kD) of UBF/UBTF (Panels A,B). This band is specifically immunoprecipitated from K562 lysates by each antibody (Panels C,D). Based on this, SAB1404509 and sc-13125 are validated by this criterion.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
UBTF (Homo sapiens)
Method: immunoprecipitation
not reviewed
Caption
Because the overlap of the top 40% of peaks from datasets derived from antibodies sc-13125 and SAB1404509 (compared using standard ENCODE scoring methods) exceeds the recommended threshold of 80%, both antibodies meet this criterion for validation.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
UBTF (Mus musculus)
Method: immunoprecipitation
not reviewed
Caption
Immunoprecipitation of CH12 and MEL nuclear extracts using anti-UBF (sc-13125) efficiently enriches a protein of molecular weight of UBF (~97 kD).
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
UBTF (Mus musculus)
Method: ChIP-seq comparison
not reviewed
Caption
sc-13125 is validated for human cell lines using comparison of ChIP-Seq data obtained using two different antibodies against UBTF. See validation documents for human cell lines for details.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
UBTF (Homo sapiens)
GM12878K562HeLa-S3HepG2
Method: immunoblot
Attachment from submitter
compliant
Caption
Western blot analysis of nuclear lysates prepared from multiple cells lines loaded in the order: GM12878, K562, HeLaS3, HepG2 using the antibody sc-13125. Molecular Weight: 97.0
Reviewer comment
No visible band in the lane
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
UBTF (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody sc-13125. Lane 1: input nuclear lysate. Lane 2: material immunoprecipitated with antibody. Lane 3: material immunoprecipitated using control IgG. Marked bands were excised from gel and subjected to analysis by mass spectrometry. Target molecular weight: 97.0.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
UBTF (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using the antibody sc-13125, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGEBis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was stained by ColloidialCoomassie G-250 stain, gel fragments corresponding to the bands indicated were excised. Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an Orbitrap Elite mass spectrometer (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Peptide false discovery rate < 1%, 2 unique peptides per protein minimum, mass error < 10 ppm).
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford