1. Antibodies
  2. Homo sapiens
  3. COMMD3-BMI1 + BMI1

ENCAB000BDV

Antibody against Homo sapiens BMI1

Homo sapiens
GM12878, K562, HeLa-S3, HepG2, IMR-90, A549, heart, endothelial cell of umbilical vein, MCF-7, HEK293T
characterized to standards
Homo sapiens
H1
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
Attachment from submitters
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
Attachment from submitters
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.jpg
BMI1 (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 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
Attachment from submitters
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
Attachment from submitters
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
Method: immunoblot
Attachment from submitter
Attachment from submitters
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
Attachment from submitters
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
Attachment from submitters
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
 expt1032_4.jpg