ENCAB000ADW

Alternate accession: ENCAB000APQ

Antibody against Homo sapiens H3K4me1, Mus musculus H3K4me1

Homo sapiens
any cell type or tissue
not characterized to standards
Bos taurus
any cell type or tissue
Status
released
Source (vendor)
Abcam
Product ID
ab8895
Lot ID
383111
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
Antigen description
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, mono methylated at K4.
Aliases
bradley-bernstein:PchAb 85-V

Characterizations

H3K4me1 (Homo sapiens)
thymus
Method: immunoblot
compliant
Submitter comment
This western blot demonstrates the antibody recognizes a protein of the molecular weight consistent with human histone H3
Reviewer comment
Though this western blot was carried out with calf biosamples, the ENCODE Antibody Review Panel has reviewed this characterization and allowed this lot to be exempted for use in human biosamples on November 11, 2016.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
H3K4me1 (Homo sapiens)
Method: dot blot assay
not compliant
Caption
Suitably modified peptides modeled on histone tails were obtained from Abcam. Each peptide is added to Bio-Dot apparatus at 0.01 and 0.1 ug / 50uL total volume (BioRad) Let sit on rocker 60 minutes. Pull through with vacuum, wash once with 100uL TBS and twice with 200uL TBSTw. Remove membrane from Bio-Dot and transfer to 10mL Pierce SuperBlock in tray. Place on rocker for 30 minutes. Dump off SuperBlock.
Reviewer comment
This antibody was found to be not compliant dot blot not demonstrating a 10 fold higher peak at the H3K4me1 mark.
Submitted by
Bradley Bernstein
Lab
Bradley Bernstein, Broad
H3K4me1 (Mus musculus)
Method: dot blot assay
not reviewed
Caption
Dr. Brad Bernstein and his colleagues at the Broad Institute have already validated this and several other antibodies directed against specific histone modifications. They spotted synthetic peptides containing one of about 20 histone modifications on a blot, in two different concentrations. The blot was then allowed to react with the antibody, and the antigen-antibody complexes were visualized and quantified. In each case, the antibody showed strong specificity. This is far better than showing a single band on a Western blot, since all the modifications we examine are on histone H3, and they all will show the H3 band. The Western blot will not demonstrate specificity for a particular modification, whereas the dot blot does. The relevant document for H3K4me1 is http://hgwdev.cse.ucsc.edu/ENCODE/validation/antibodies/ human_H3K4me1_validation_Bernstein.pdf Our validation point for mouse is that there is nothing species-specific about the existing validations. Synthetic peptides were used on the blot, and the assay was for specific reaction with the antibody. The peptides on the blot were not species-specific because HUMAN AND MOUSE HISTONE H3 ARE IDENTICAL IN THE RELEVANT REGIONS. Human and mouse H3 differ at only one position, amino acid 97, where a Cys in human is replaced by a Ser in mouse. There are NO differences in the relevant region, which is the N-terminal 36 amino acids.
Submitted by
Ross Hardison
Lab
Ross Hardison, PennState