1. Antibodies
  2. Homo sapiens
  3. SMARCC1

ENCAB000AEB

Antibody against Homo sapiens SMARCC1

Homo sapiens
at least one cell type or tissue
awaiting characterization
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-10756
Lot ID
unknown
Characterized targets
SMARCC1 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Antigen description
Raised against amino acids 998-1073 of BAF155 of human origin.

Characterizations

SMARCC1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
not reviewed
Caption
Methodology HeLaS3 whole cell lysates were immunoprecipitated using sc10756, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGE Bis-Tris Gel) and separated with an invitrogen NuPAGE electrophoresis system. The gel was silver-stained, gel fragments corresponding to the bands indicated were excised and destained using the SilverSNAP Stain for Mass Spectrometry (Pierce). Then proteins were trypsinized using the in-gel digestion method. The samples were subjected to nanoflow chromatography using the nanoAcquity UPLC system (Waters Inc.) prior to introduction into the mass spectrometer for further analysis. Mass spectrometry was performed on a hybrid ion trap LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific) in positive electrospray ionization (ESI) mode. The spectra was acquired in a data dependent fashion consisting of full mass spectrum scan (300-€“2000 m/z) followed by MS/MS scan of the 3 most abundant parent ions. For the full scan in the orbitrap the automatic gain control (AGC) was set to 16106 and the resolving power for 400 m/z of 30,000. The MS/MS scans were done using the ion trap part of the mass spectrometer at a normalized collision energy of 24 V. Dynamic exclusion time was set to 100 s to avoid loss of MS/MS spectral information due to repeated sampling of the most abundant peaks. Sequence data from MS/MS spectra was processed using the SEQUEST database search algorithm (Thermo Fisher Scientific). The resulting protein identifications were brought into the Scaffold visualization software (Proteome Software) where the information was further refined resulting in improved protein id conformation. Scaffold search criteria were set at 98% probability and required at least 2 unique peptides per id. Mass spectrometry identifies BAF155/SMARCC1 as the major component of the material immunoprecipitated with antibody sc-10756 and excised from the bands indicated in the figure. A more detailed presentation of this data has been published (Euskirchen et.al., (2011) PLoS Genetics, 7(3):1-20), where an analysis of all associated proteins in presented. This work confirms previous observations that many components of the SWI/SNF complex are associated with BAF155/SMARCC1, including three additional members, BAF170/SMARCC2, Brg1/SMARCA4, and Ini1/SMARCB1, for which ENCODE ChIP-seq datasets are currently available.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
SMARCC1 (Homo sapiens)
Method: immunoprecipitation
Attachment PDF Icon
not reviewed
Caption
A band consistent with the commonly observed mobility of BAF155/SMARCC1 (~155kD) is immunoprecipitated from HeLaS3 nuclear lysate using sc-10756. An additional band of lesser abundance is observed at ~120kD. Mass spectrometry confirms that both bands contains BAF155 in high abundance (see validation #2).
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford