ENCAB494QXU

Antibody against Homo sapiens H3F3A

Homo sapiens
any cell type or tissue
characterized to standards with exemption
Status
released
Source (vendor)
Millipore
Product ID
09-838
Lot ID
2273984
Characterized targets
H3F3A (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Antigen description
H3.3 AIG
Aliases
bradley-bernstein:PchAb 1071
External resources

Characterizations

H3F3A (Homo sapiens)
Method: dot blot assay
Attachment from submitter
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
H3F3A (Homo sapiens)
K562HEK293
Method: immunoblot
Attachment from submitter
compliant
Caption
6ug of unmodified Recombinant Histone H3 variants H3.1, H3.2 and H3.3 and 2*10^5 cells equivalent of HEK and k562 Nuclear lysates 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 H3 (Abcam ab1791), band of expected size (~17kDa) visualized in all histone H3 variants, at similar intensities.Panel B: anti H3.3 (Millipore 09-838) Band of expected size (~17kDa) visualized in Histone H3.3 variant only, and cannot be detected in other H3 variants. Panel C: a single band of expected size was detected in both nuclear lysates.The antibody seems to be specific since no reactivity with non histone proteins or other histone H3 variants was detected
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad