ENCAB000BDV

Antibody against Homo sapiens BMI1

Homo sapiens
A549, GM12878, endothelial cell of umbilical vein, heart, HeLa-S3, IMR-90, K562, MCF-7, HepG2, HEK293T
characterized to standards
Homo sapiens
H1-hESC
not characterized to standards
Status
released
Source (vendor)
Cell Signaling
Product ID
6964S
Lot ID
1
Characterized targets
BMI1 (Homo sapiens)
Host
rabbit
Clonality
monoclonal
Purification
affinity
Isotype
IgG
Antigen description
The carboxy terminus of human Bmi1 protein.
Aliases
bradley-bernstein:PchAb 774
External resources

Characterizations

BMI1 (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T using the antibody 6964S. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 41, 43.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
BMI1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation of Bmi1 from K562 cells using 6964S. Lane 1: input nuclear lysate. Lane 2: material immunoprecipitated with 6964S. Lane 3: material immunoprecipitated using control IgG. Band A was excised from gel and subject to analysis by mass spectrometry. Expected band size is 43 kDa.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
Download
BMI1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
compliant
Caption
IP followed by mass spectrometry: Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using 6964S, 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
For this factor, we found DNA-binding proteins SMARCE1, DPF2, RING1, RNF2 and ACTL6A have more peptides than targeted protein Bmi1. We would like to explain as follows: Bmi1 has unique interaction with DPF2, and DPF2 also has interaction with SMARCE1, which one can find here: http://thebiogrid.org/111909/summary/homo-sapiens/dpf2.html SMARCE1 has interactions with RING1, RNF2 and ACTL6A, it can be found here: http://thebiogrid.org/112489/summary/homo-sapiens/smarce1.html
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
BMI1 (Homo sapiens)
GM12878K562HeLa-S3HepG2IMR-90A549heart
Method: immunoblot
Attachment from submitter
compliant
Caption
Western blot analysis of nuclear lysates prepared from multiple cells lines loaded in the order : GM12878, K562, HelaS3, HepG2, IMR90, A549 and Heart using the antibody 6964S. Molecular mass: 36949 Da
Submitted by
Trupti Kawli
Lab
Michael Snyder, Stanford
BMI1 (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 6964S .The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG. Expected size ~37 kDa.
Submitted by
Trupti Kawli
Lab
Michael Snyder, Stanford
BMI1 (Homo sapiens)
K562A549H1-hESC
Method: immunoblot
Attachment from submitter
compliant
Caption
Nuclear lysates from K562 (10ug), A549 (10ug) and H1 (10ug) were loaded into a 4-12% Bis-Tris gel in 1X MOPS running buffer. After separation, the samples were transferred to a nitrocellulose membrane using iblot. Membrane was blocked for an hour in room temperature, with 5% BSA in TBS-T 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. Only one band at the expected size (45kDa) was detected.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
BMI1 (Homo sapiens)
HepG2GM12878endothelial cell of umbilical vein
Method: immunoblot
Attachment from submitter
compliant
Caption
Nuclear lysates from GM12878 (10ug),HUVEC (10ug), HepG2 (10ug), were loaded into a 4-12% Bis-Tris gel in 1X MES running buffer. After separation, the samples were transferred to a nitrocellulose membrane using iblot. Membrane was blocked for an hour in room temperature, with 5% BSA in TBS-T 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. The only band detected was of the expected size (~40kDa). **Molecular mass: 36949 Da**
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
BMI1 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody 6964S. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
Download