ENCAB000AKO

Antibody against Homo sapiens SAP30

Homo sapiens
any cell type or tissue
partially characterized
Homo sapiens
HepG2, K562, SK-N-SH, GM12878
not characterized to standards
Status
released
Source (vendor)
Active Motif
Product ID
39731
Lot ID
4710001
Characterized targets
SAP30 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
crude
Antigen description
Raised against recombinant full length human SAP30 protein.
Aliases
bradley-bernstein:PchAb 164-V

Characterizations

SAP30 (Homo sapiens)
Method: ChIP-string comparison
not reviewed
Caption
Antibody was validated by use of correlation analysis of ChIP-String and ChIP-Seq data
Submitted by
Bradley Bernstein
Lab
Bradley Bernstein, Broad
SAP30 (Homo sapiens)
Method: immunoblot
not reviewed
Submitted by
Bradley Bernstein
Lab
Bradley Bernstein, Broad
SAP30 (Homo sapiens)
K562HepG2GM12878SK-N-SH
Method: immunoblot
Attachment from submitter
not compliant
Reviewer comment
KCO: None of the major bands shown on the gel are within 20% of the expected band size (30 kDa).
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
SAP30 (Homo sapiens)
K562
Method: immunoblot
Attachment from submitter
not compliant
Caption
0.25ug unmodified Recombinant Histone H3 (Millipore 14-494) and 30ug of K562 Whole Cell Lysate (Abcam, ab7911) were resolved by electrophoresis on a 4-12% acrylamide gel. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen iBlot system. Membrane was blocked for an hour in room temperature, with 5% nonfat dry milk and blotted with primary antibody in the appropriate concentration over night at 4c. Membrane was washed and blotted with secondary HRP-conjugated antibody. Detection was made with Optiblot ECL Detect Kit (ab133406) for 2 min. Panel A: anti Histone H3 (Abcam ab1791), band of expected size (~17kDa) visualized in both unmodified recombinant histone H3 and WCL, at similar intensities.Panel B: Band of expected size (~17kDa) visualized in WCL, and cannot be detected in unmodified recombinant Histone H3. Antibody seems to be specific since no reactivity with nonhistone proteins or unmodified histones was detected
Reviewer comment
KCO: Image (and caption) neither provides the cell line used, nor the precise expected band size value (based on Antibody Characterization Standards, August 2015)
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
SAP30 (Homo sapiens)
Method: ChIP-seq comparison
compliant
Caption
This validation relies on the use of antibodies to different members of a known complex, in K562 cells, and the demonstration that highly similar patterns of enrichment are obtained with each antibody. The second track shown used an antibody to HDAC1 (PchAb 134-V), another member of the Sin3a complex. Overall correlation score: SAP30 vs. HDAC1: 0.9303
Reviewer comment
Currently under "pending dcc review" because the document needs to include the accession of the antibody used for comparison, rather than just an internal identifier.
Submitted by
Kathrina Onate
Lab
Bradley Bernstein, Broad