1. Antibodies
  2. Homo sapiens
  3. MCM7

ENCAB658SAJ

Antibody against Homo sapiens MCM7

Homo sapiens
K562
characterized to standards with exemption
Status
released
Source (vendor)
Active Motif
Product ID
13/61725
Lot ID
17015001
Characterized targets
MCM7 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Isotype
IgG
Aliases
michael-snyder:AS-829

Characterizations

MCM7 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562 using the antibody 13/61725. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 81.308.
Reviewer comment
Doublet near expected size but marked band > 50% of total signal
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
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 K562-MCM7-1.jpg
MCM7 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
exempt from standards
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using the antibody 13/61725, 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
MCM7 peptides were detectable in the IP, as well as, in the IgG lane. As the number of unique MCM7 peptides in the IP lane was higher than for other proteins, this mass spec analysis was exempt from the standards by the ENCODE antibody review panel. 2 other proteins with high peptide counts, MCM6 and MCM4, are part of of a complex with MCM7 (http://www.genecards.org/cgi-bin/carddisp.pl?gene=mcm7).
Reviewer comment
Although MCM7 is not in the top 25 ranked peptides by enrichment over IP, MCM6 and other components of the same complex are. On 10/21/16, the ENCODE Antibody Review Panel exempted this characterization.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
MCM7 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody 13/61725. 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: 81.308.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford