1. Antibodies
  2. Homo sapiens
  3. YBX1

ENCAB493UWX

Antibody against Homo sapiens YBX1

Homo sapiens
K562
characterized to standards
Homo sapiens
MCF-7, GM12878, HepG2, HEK293T
characterized to standards with exemption
Homo sapiens
any cell type or tissue
partially characterized
Status
released
Source (vendor)
Bethyl Labs
Product ID
A303-230A
Lot ID
1
Characterized targets
YBX1 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
External resources

Characterizations

YBX1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody A303-230A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Reviewer comment
signal is in the IgG
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
YBX1 (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 A303-230A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 35.924
Submitter comment
We'd like an exemption for this characterization.
Reviewer comment
Not within 20% of expected size, but on 8/5/16, the size discrepancy was exempted by the antibody review panel because the mass spec in K562 detects the TF in a similarly sized band.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
YBX1 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody A303-230A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 35.924
Submitter comment
Please exempt this characterization.
Reviewer comment
Not within 20% of expected size, but on 8/5/16, the size discrepancy was exempted by the antibody review panel because the mass spec in K562 detects the TF in a similarly sized band.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
YBX1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody A303-230A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
YBX1 (Homo sapiens)
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not submitted for review by lab
Caption
IP-WB analysis of K562 whole cell lysate using YBX1 specific antibody. Lane 1 is 2.5% of 0.5mg input lysate, lane 2 is 2.5% of supernatant after immunoprecipitation and Lane 3 is 50% of IP enrichment using rabbit polyclonal anti-YB1. This antibody did not meet our primary validation criteria using our standard IP protocol in the indicated cell type.
Submitted by
Balaji Sundararaman
Lab
Gene Yeo, UCSD
YBX1 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878, using the antibody A303-230A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 35.924
Submitter comment
Please exempt this characterization.
Reviewer comment
Not within 20% of expected size, but on 8/5/16, the size discrepancy was exempted by the antibody review panel because the mass spec in K562 detects the TF in a similarly sized band.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
YBX1 (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody A303-230A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 35.924
Submitter comment
The TF was detected in a similarly sized band in the K562 IP-MS characterization.
Reviewer comment
Not within 20% of expected size, but on 8/5/16, the size discrepancy was exempted by the antibody review panel because the mass spec in K562 detects the TF in a similarly sized band.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
YBX1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody A303-230A. 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: 35.924.
Submitter comment
--
Reviewer comment
Band is not within 20% of expected size, but rescued by mass spectrometry analysis
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
Download
 YBX1.jpeg
YBX1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using the antibody A303-230A, 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
None of the detected proteins are DNA-binding proteins
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford