1. Antibodies
  2. Homo sapiens
  3. TBX3

ENCAB320XLX

Antibody against Homo sapiens TBX3

Homo sapiens
HepG2
characterized to standards with exemption
Status
released
Source (vendor)
Bethyl Labs
Product ID
A303-098A
Lot ID
1
Characterized targets
TBX3 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1033
External resources

Characterizations

TBX3 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2, using the antibody A303-098A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Reviewer comment
Multiple bands detected and indicated size not at least 50% of the signal. Could potentially be rescued with a knockdown characterization.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
TBX3 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line HepG2 using the antibody A303-098A. 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: 79.389.
Submitter comment
We analyzed multiple immunoreactive bands by mass-spec.
Reviewer comment
Multiple bands but mass spec analysis showed the target TF to be highest ranked and not in IgG with no other site specific TFs above the target.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
TBX3 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
exempt from standards
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from HepG2 nuclear cell lysates using the antibody A303-098A, 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
We can not explain the higher ranked TFs.
Reviewer comment
PF: I’m not worried about YLPM1 and TAF15 because there is no evidence that these are likely to be site-specific TFs. But the issue is ZNF687- this one has more peptides than the target TF but neither are detected in the control IgG. We can’t say that it is not compliant since there are no peptides in the control IgG.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford