ENCAB201MQH
Antibody against Homo sapiens ILF3
Homo sapiens
Jurkat, Clone E6-1
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)
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
- Grant
- U54HG006996
- Download
- ILF3_612155 final.pdf
ILF3 (Homo sapiens)
K562
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
- Grant
- U54HG006996
- Download
- MS1047_6_ILF3-612155.JPG
ILF3 (Homo sapiens)
Jurkat, Clone E6-1
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
- Grant
- U54HG006996
- Download
- NF90_Ab_validation_Jurkat.jpeg