1. Antibodies
  2. Homo sapiens
  3. H2AFZ

ENCAB000ASY

Antibody against Homo sapiens H2AFZ

Homo sapiens
any cell type or tissue
characterized to standards with exemption
Status
released
Source (vendor)
Millipore
Product ID
07-594
Lot ID
DAM1540736
Characterized targets
H2AFZ (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
Protein A/G
Isotype
serum
Antigen description
Raised against a peptide corresponding to the C-terminus of H2A.Z
Aliases
bradley-bernstein:PchAb 64-V

Characterizations

H2AFZ (Homo sapiens)
K562HEK293
Method: immunoblot
Attachment from submitter
Attachment from submitters
compliant
Caption
0.25ug unmodified Recombinant Histone H2A (NEB M2502S) and 1.5*10^6 cells equivalent of HEK Nuclear lysate were resolved by electrophoresis on a 4-12% acrylamide gel. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen iBlot system. Membrane was blocked for an hour in room temperature, with 5% nonfat dry milk 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. Panel A: anti Histone H2A (Abcam ab18255), band of expected size (~14kDa) visualized in both unmodified recombinant histone H2A and NL, at similar intensities.Panel B: Band of expected size (~14kDa) visualized in NL, and cannot be detected in unmodified recombinant H2A. The antibody seems to be specific since no reactivity with nonhistone proteins or unmodified histones was detected
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
H2AFZ (Homo sapiens)
Method: dot blot assay
Attachment from submitter
Attachment from submitters
exempt from standards
Submitter comment
The validations rely on western blots. We believe that these blots satisfy primary and secondary validation criteria. The western blots show that each antibody binds an epitope of the expected size (primary validation) and does not bind to a recombinant source of the corresponding canonical histone, and is thus specific to the non-canonical histone variant (secondary validation). Because these are antibodies to histone variants, rather than post - translational modifications of histones, we do not use our array of modified peptides.
Reviewer comment
I think that these are very believable validation methods. I would approve the antibodies that are shown in the file that was attached and would like to ask Chuck to add a paragraph to the Antibody document that deals with histone variants, using the methods he has described.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
H2AFZ (Homo sapiens)
Method: immunoblot
Attachment from submitter
Attachment from submitters
not reviewed
Submitter comment
This is from the company
Submitted by
Bradley Bernstein
Lab
Bradley Bernstein, Broad
H2AFZ (Homo sapiens)
Method: ChIP-seq comparison
Attachment from submitter
Attachment from submitters
not reviewed
Caption
Antibody was validated by analysis of ChIP-Seq data comparing tracks derived using different lots of the antibody in ChIPs performed on different cell types.
Submitted by
Bradley Bernstein
Lab
Bradley Bernstein, Broad