ENCAB506UYG

Antibody against Homo sapiens TCF12

Homo sapiens
GM12878, K562
characterized to standards with exemption
Status
released
Source (vendor)
Bethyl Labs
Product ID
A300-754A
Lot ID
1
Characterized targets
TCF12 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1445
External resources

Characterizations

TCF12 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody A300-754A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 72.965
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
TCF12 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody A300-754A. 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: 72.965.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
TCF12 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
exempt from standards
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using the antibody A300-754A, 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).
Submitter comment
NUP155 is not DNA binding protein. DLAT is located in mitochondrion. SMARCA5 was also detected in MS of TCF12 (ENCAB521XDP).
Reviewer comment
TCF12 is not the highest ranked TF in the IP-MS analysis but SMARCA5, which is ranked higher is also detected in an independent IP-MS analysis of another TCF12 antibody (ENCAB521XDP) so it's possible that is due to a novel interaction not yet described in the literature. This case is permitted by the standards document.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
TCF12 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878 using the antibody A300-754A. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 72.965.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford