1. Antibodies
  2. Homo sapiens
  3. NBN

ENCAB000BMB

Alternate accession: ENCAB551CQT

Antibody against Homo sapiens NBN

Homo sapiens
HepG2
characterized to standards
Homo sapiens
GM12878, K562, HeLa-S3, MCF-7, liver
not characterized to standards
Status
released
Source (vendor)
Sigma
Product ID
HPA001429
Lot ID
R07264
Characterized targets
NBN (Homo sapiens)
Lot ID aliases
A97300
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
Antigen description
Nibrin recombinant protein epitope signature tag (PrEST)
Antigen sequence
DTFSDEAVPESSKISQENEIGKKRELKEDSLWSAKEISNNDKLQDDSEMLPKKLLLTEFRSLVIKNSTSRNPSGINDDYGQLKNFKKFKKVTYPGAGKLPHIIGGSDLIAHHARKNTELEEWLRQEMEVQNQHAKEESLADDL
Aliases
michael-snyder:AS-1127
External resources

Characterizations

NBN (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line HepG2 using the antibody HPA001429_(ENCAB000BMB). 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: 85.0.
Reviewer comment
Multiple immunoreactive bands but mass spec shows target TF in both marked regions.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
NBN (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from HepG2 nuclear cell lysates using the antibody HPA001429_(ENCAB000BMB), 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).
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
NBN (Homo sapiens)
GM12878K562HepG2HeLa-S3MCF-7liver
Method: immunoblot
Attachment from submitter
Attachment from submitters
not compliant
Caption
Western blot analysis of nuclear lysates prepared from multiple cells lines loaded in the order : GM12878, K562, HepG2, HelaS3, MCF7 and Liver using the antibody HPA001429. Molecular mass: 84959 Da
Reviewer comment
Band is not 50% of overall signal
Submitted by
Trupti Kawli
Lab
Michael Snyder, Stanford
NBN (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
b) Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2, using the antibody HPA001429 .The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG. Expected size ~85 kDa.
Reviewer comment
Band is not 50% of total signal
Submitted by
Trupti Kawli
Lab
Michael Snyder, Stanford