ENCAB543JMP

Antibody against Homo sapiens ZBTB2

Homo sapiens
HepG2, K562, MCF-7
characterized to standards with exemption
Status
released
Source (vendor)
Bethyl Labs
Product ID
A303-262A
Lot ID
1
Characterized targets
ZBTB2 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1469
External resources

Characterizations

ZBTB2 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody A303-262A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 57.337
Reviewer comment
Band is too high in size, beyond the allowable 20% of expected size.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
ZBTB2 (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 A303-262A. 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: 57.337.
Submitter comment
We detected ZBTB2 by mass spec in the marked immunoreactive band.
Reviewer comment
Runs higher than expected size but ZBTB2 is detected by accompanying mass spec.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZBTB2 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2 using the antibody A303-262A. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 57.337.
Submitter comment
Similar in size to K562 IP-MS.
Reviewer comment
Runs higher than expected size but banding pattern is consistent across cell types and was detected in similarly-sized band by mass spec in K562.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZBTB2 (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 A303-262A, 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
RMDN3, NCL, UBC, ALB, C3 are not TFs. FUS, HSPA1A and LAS1L are RNA-binding proteins. Both MTA1 and HDAC1 have interactions with ZBTB2, https://thebiogrid.org/121667/summary/homo-sapiens/zbtb2.html. Also, MTA1 has interaction with HDAC1, 17 publications on biogrid, https://thebiogrid.org/109315/summary/homo-sapiens/hdac1.html. MTA2, CHD4, GATAD2B, ZMYND8, HDAC2, GATAD2A, MTA3 and DDX17 have interactions with HDAC1, https://thebiogrid.org/109315/summary/homo-sapiens/hdac1.html. Both NRF1 and HDAC1 have interactions with HCFC1 and Jun, https://thebiogrid.org/109315/summary/homo-sapiens/hdac1.html and https://thebiogrid.org/110851/summary/homo-sapiens/nfe2l1.html. Also, HCFC1 and HDAC1 are in one complex, http://www.genecards.org/cgi-bin/carddisp.pl?gene=HCFC1&keywords=hcfc1. HCFC1 (209KD) and Jun (36KD) are not in the gel slice with NRF1 (54KD), HDAC1 (55KD) and ZBTB2. PDCD11 is a NFKB1 binding protein, NFKB1 and HDAC1 have interactions, 6 publications on biogrid, https://thebiogrid.org/109315/summary/homo-sapiens/hdac1.html. ZNF800 has interactions with MTA2, https://thebiogrid.org/114652/summary/homo-sapiens/mta2.html. NKRF is NF-KappaB Repressing Factor, NFKB1 and HDAC1 have interactions, 6 publications on biogrid, https://thebiogrid.org/109315/summary/homo-sapiens/hdac1.html.
Reviewer comment
ZBTB2 is not the top ranked TF in the peptide list but lab has provided references for interactions with ZBTB2 and interacting partners of ZBTB2.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ZBTB2 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7 using the antibody A303-262A. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 57.337.
Submitter comment
Similar in size to K562 IP-MS.
Reviewer comment
Runs higher than expected size but banding pattern is consistent across cell types and was detected in similarly-sized band by mass spec in K562.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford