ENCAB939ONI

Antibody against Homo sapiens GATAD2B

Homo sapiens
MCF-7, GM12878, K562, HepG2, HEK293T
characterized to standards
Status
released
Source (vendor)
Bethyl Labs
Product ID
A301-281A
Lot ID
1
Characterized targets
GATAD2B (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1612
External resources

Characterizations

GATAD2B (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody A301-281A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 65.261
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
GATAD2B (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2, using the antibody A301-281A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 65.261
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
GATAD2B (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody A301-281A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 65.261
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
GATAD2B (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878, using the antibody A301-281A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 65.261
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
GATAD2B (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody A301-281A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 65.261
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
GATAD2B (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 A301-281A, 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
HDAC1 and FUS peptides were detected. HDAC1 has been shown to interact with GATAD2B (http://thebiogrid.org/121529/summary/homo-sapiens/gatad2b.html, 6 publications). No direct interaction between FUS and GATAD2B has been shown so far but HDAC1’s Mass-Spec results (ENCAB447WPM) detected a indirect interaction with FUS via SPI1. TPM4 is actin-binding protein. HNRNPA2B1 is RNA binding protein.
Reviewer comment
Submitters showed that FUS1 interacts indirectly with HDAC1 via SPI1 in their IP-MS data and HDAC1 itself interacts with GATAD2B. The standards stipulate that if another TF is ranked higher than the target TF, that if it it has a known interaction with another protein known to interact with the TF then it can be deemed compliant.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
GATAD2B (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody A301-281A. 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: 65.261.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford