ENCAB201MQH

Antibody against Homo sapiens ILF3

Homo sapiens
Jurkat clone E61
characterized to standards
Homo sapiens
K562
characterized to standards with exemption
Status
released
Source (vendor)
BD Biosciences
Product ID
612155
Lot ID
6104880
Characterized targets
ILF3 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Purification
affinity
Isotype
IgG1
Antigen description
Human DRBP76 aa. 592-695
Aliases
michael-snyder:AS-1669

Characterizations

ILF3 (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 612155, 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
In band A, none of the proteins ranked equally have sequence-specific TF activity. In band B, none of the proteins ranked above are sequence-specific TFs. Among the equally ranked proteins in band B is HMGA1 which is a sequence-specific TF. ILF3 and HMGA1 have not been shown to interact, however they have several interactors in common, such as CDK2, CUL7, EGFR, HNRNPA1, HNRNPL, NCL, and PRMT6 (https://thebiogrid.org/109822/summary/homo-sapiens/ilf3.html and https://thebiogrid.org/109402/summary/homo-sapiens/hmga1.html).
Submitted by
Jessika Adrian
Lab
Michael Snyder, Stanford
ILF3 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody 612155. 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: 95.338.
Submitter comment
ILF3 protein has 7 isoforms produced by alternative splicing. Most isoforms have a molecular weight of around 75 and 95 kD (source: http://www.uniprot.org/uniprot/Q12906).
Reviewer comment
Multiple immunoreactive bands seen but subsequent mass-spec analysis detects the TF of interest in the analyzed bands.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ILF3 (Homo sapiens)
Jurkat clone E61
Method: immunoprecipitation
Attachment from submitter
compliant
Submitter comment
Immunoprecipitation was performed on nuclear extracts from Jurkat cells using the antibody BD 612154/612155 (same product ID, different volumes). The image shows western blot analysis of input, immunoprecipitate, flowthrough, and mock immunoprecipitate using IgG. Target molecular weight: 90 kD.
Submitted by
Jessika Adrian
Lab
Michael Snyder, Stanford