Histone H3 antibody (mAb)

RRID: AB_2687473
Clone: 1B1-B2
Catalog No: 61475 Format: 100 µg Discontinued
Catalog No: 61976 Format: 50 µg Discontinued
Catalog No: 61476 Format: 10 µg Discontinued

Antibody Type:
Monoclonal
Isotype:
IgG3
Purification:
Protein A Chromatography
Host:
Mouse
Molecular Weight:
17 kDa
Reactivity:
Human, Wide Range Predicted

Applications

ChIP Validated ChIP-Seq Validated Western Blot Validated

Application Notes


This product has been discontinued
Validated Applications:
ChIP-Seq: 4 µg per ChIP
ChIP: 5 µg per ChIP
WB: 1 - 4 µg/ml dilution

Published Applications:
WB
IP

Immunogen

This antibody was raised against a peptide within the C-terminal region of human histone H3.

Buffer

Purified IgG in 100mM sodium phosphate pH 8.0, 90mM trisodium citrate, 430mM sodium chloride.
If crystals are noted, warm to room temperature and mix gently before use.

References

Dal-Pra et. al., Circ Res. 2017; 120:1403-1413. (http://circres.ahajournals.org/content/120/9/1403)
Connelly et. al., Yale J. Biol Med. 2016; Dec; 89(4): 431-440. (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5168823/)

 

Histone H3 antibody (mAb) tested by ChIP-Seq.Chromatin immunoprecipitation (ChIP) was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 30 µg of chromatin from Farage human lymph-node derived B cells and 5 µl Histone H3 antibody. ChIP DNA was sequenced on the Illumina HiSeq and 16.7 million sequence tags were mapped to identify Histone H3 binding sites.

H3 antibody (mAb) tested by ChIP.
ChIP was performed using the ChIP-IT® High Sensitivity Kit (Cat. No. 53040) with 15 µg of chromatin from both HeLa and K562 cells and 4 µg H3 antibody. ChIP DNA was used in qPCR with the gene-specific primer as indicated. Data are presented as Binding Events Detected per 1000 Cells using Active Motif's Epigenetic Services normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.

Histone H3 antibody (mAb) (Clone 1B1-B2) tested by Western blot.
Histone H3 antibody detection by Western blot. The analysis was performed using 20 µg of HeLa nuclear extract and Histone H3 antibody at a 0.25 µg/ml dilution.

Background

Histone H3 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 147 base pairs of DNA wrapped around an octamer of core histone proteins (two each of Histone H2A, Histone H2B, Histone H3 and Histone H4). Chromatin is subject to a variety of chemical modifications, including post-translational modifications of the histone proteins and the methylation of cytosine residues in the DNA. Reported histone modifications include acetylation, methylation, phosphorylation, ubiquitylation, glycosylation, ADP-ribosylation, carbonylation and SUMOylation; these modifications play a major role in regulating gene expression.

Positive Control

Active Motif's HeLa nuclear extract (Catalog No. 36010) can be used as a positive control for Histone H3 antibody.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Histone H3 antibody (mAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting