ENCAB521XDP

Antibody against Homo sapiens TCF12

Homo sapiens
HepG2, K562
characterized to standards with exemption
Status
released
Source (vendor)
Bethyl Labs
Product ID
A300-755A
Lot ID
1
Characterized targets
TCF12 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1446
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 A300-755A. 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 A300-755A. 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 A300-755A, 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
All of ILF3, SMARCA5, NOP56, FUS, NOP58, EFTUD2 and NAT10were detected in MS of TCF12 (ENCAB506UYG).
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 (ENCAB506UYG) 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)
HepG2
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2 using the antibody A300-755A. 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