ENCAB000BOG

Antibody against Homo sapiens SOX6

Homo sapiens
HEK293T, K562
characterized to standards
Homo sapiens
HepG2
characterized to standards with exemption
Homo sapiens
GM12878, MCF-7, liver
not characterized to standards
Status
released
Source (vendor)
Sigma
Product ID
HPA001923
Lot ID
A81784
Characterized targets
SOX6 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
Antigen description
Transcription factor SOX-6 recombinant protein epitope signature tag (PrEST)
Antigen sequence
TYKPGDNYPVQFIPSTMAAAAASGLSPLQLQQLYAAQLASMQVSPGAKMPSTPQPPNTAGTVSPTGIKNEKRGTSPVTQVKDEAAAQPLNLSSRPKTAEPVKSPTS
External resources

Characterizations

SOX6 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2, using the antibody HPA001923. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 92.0
Reviewer comment
Band near 37kD size is more prominent than the one at the expected size.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
SOX6 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody HPA001923. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Reviewer comment
Multiple bands, marked band not >50% and would possibly need siRNA knockdown followup to be exempted
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
Download
SOX6 (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody HPA001923. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 92.0
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
SOX6 (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 HPA001923. 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: 92.0.
Submitter comment
We'll analyze all immunoreactive bands by mass-spec.
Reviewer comment
Multiple immunoreactive bands, with the major band not at the expected size. Subsequent IP-MS did reveal the target factor in the band around expected size (A) and a putative interactor in the major band (B)
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
SOX6 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from HepG2 nuclear cell lysates using the antibody HPA001923, 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
SOX6 has been shown to interact with SOX5 (https://thebiogrid.org/120714/summary/homo-sapiens/sox6.html). The SOX5 protein appears to have different isoforms, some of which are 86 and 41 kDa. In the higher band A, both proteins are ranked with the the same peptide count. In the lower band B, only SOX5 was detected. No other sequence specific TFs were ranked equally or above SOX6/SOX5 in band A and B. No sequence specific TF was detected in band C.
Reviewer comment
SOX6 is the highest ranked TF detected in band A but was not detected in bands B or C. SOX5 is detected in band B and the submitting lab has referenced an interaction between SOX5 and SOX6 by AP-MS in the published literature.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
SOX6 (Homo sapiens)
GM12878K562HepG2liver
Method: immunoblot
Attachment from submitter
not compliant
Caption
a) Western blot analysis of nuclear lysates prepared from multiple cells lines loaded in the order: GM12878, K562, HepG2, Liver using the antibody HPA001923.
Reviewer comment
Multiple bands and most prominent band not that of expected size (~97kD - also not indicated on blot or caption).
Submitted by
Trupti Kawli
Lab
Michael Snyder, Stanford
SOX6 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
b) Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody HPA001923 .The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Submitted by
Trupti Kawli
Lab
Michael Snyder, Stanford