Histone

Histone H3K4me1 Antibody, SNAP-ChIP Certified *DISCONTINUED*

SKU: 13-0040
Pack Size: 100 µg

Product Discontinued - See 13-0057 as a recommended alternative

Type:  Mixed Monoclonal* Host:  Rabbit
Mol Wgt.:  15 kDa Appl.:  ChIP, ChIP-Seq, WB, ICC, E, L
Format:  Aff. Pur. IgG Reactivity: H, M, WR
Histone H3K4me1 Antibody, SNAP-ChIP Certified Description: This antibody meets EpiCypher’s “SNAP-ChIP® Certified” criteria for specificity and efficient target enrichment in a ChIP experiment (<20% cross-reactivity across the panel, >5% recovery of target input). Histone H3 is one of the four proteins that are present in the nucleosome, the basic repeating subunit of chromatin, consisting of 147 base pairs of DNA wrapped around an octamer of core histone proteins (H2A, H2B, H3 and H4). This antibody reacts to H3K4me1 when present alone as well as in combination with nearby acetylations (tetraAc-H3). No cross reactivity with extended acyl states (H3K9bu, H3K9cr) or other lysine acylations in the EpiCypher SNAP-ChIP K-AcylStat panel, is detected.
*Mixed Monoclonal: a pool of multiple monoclonal antibodies.

Histone H3K4me1 Antibody, SNAP-ChIP Certified Certified Immunogen: A synthetic peptide corresponding to histone H3 monomethylated at lysine 4.

Histone H3K4me1 Antibody, SNAP-ChIP Certified Certified Formulation: Protein A affinity-purified antibody (1 mg/mL) in PBS, pH 7.2 with 0.09% sodium azide

Histone H3K4me1 Antibody, SNAP-ChIP Certified Certified Storage and Stability: Stable for 1 years at -20°C from date of receipt.

Histone H3K4me1 Antibody, SNAP-ChIP Certified Certified Application Notes Recommended dilutions:
 
ChIP: 2 - 5 μg per 1x106 cells IF/ICC: 1 μg/mL WB: 0.5 - 1 μg/mL L: 0.5 - 2 μg/mL

Histone H3K4me1 Antibody, SNAP-ChIP Certified Certified References:

Grzybowski et al (2015) Mol Cell 58:886

Shah et al (2018) Mol Cell 72:162

View technical datasheet for this product. 13-0040 Datasheet

Applications Key: ChIP-Chromatin IP; E-ELISA; FACS-Flow cytometry; IF-Immunofluorescence; IHC-Immunohistochemistry; ICC-Immunocytochemistry; L-Luminex; IP-Immunoprecipitation; WB-Western Blotting

Reactivity Key: B-Bovine; Ce-C. elegans; Ch-Chicken; Dm- Drosophila; Eu-Eukaryote; H-Human; M-Mouse; Ma-Mammal; R-Rat; Sc-S.cerevesiae; Sp-S. pombe; WR-Wide Range (predicted); X-Xenopus; Z-Zebrafish

13-0040 SNAP-ChIP Data

Representative SNAP-ChIP-seq results: Cumulative histogram plot and heatmap of signal intensity depict H3K4me1 ChIP-seq data aligned to annotated transcription start sites (TSS, +/- 3.0 kb; left). Two representative genomic regions depicting H3K4me1 peak structure and distribution are shown (right). Data shown are representative of H3K4me1 ChIP antibody (EpiCypher Catalog No. 13-0040) and are not lot-specific. Native ChIP-seq was performed using K562 cells as described (Shah et al., Mol Cell 2018) with SNAP-ChIPTM K-MetStat Spike-in (Catalog No. 19-1001) nucleosome controls added prior to chromatin digestion to confirm antibody specificity and ChIP efficiency. Paired-end sequencing libraries were prepared using the NEBNext® UltraTM II DNA Library Prep Kit for Illumina®. ChIP libraries were sequenced on an Illumina® NextSeq. Sequencing reads were aligned to the human genome using Bowtie 2 (Johns Hopkins University). Bigwig files of read enrichment in binned genomic regions (signal intensity) flanking the indicated gene features were used to create a cumulative histogram plot and heatmap of signal intensity (www.basepairtech.com). Gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute) with the window size denoted (top).
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13-0040 SNAP-ChIP Data

SNAP-ChIP qPCR Data: Histone H3K4me1 antibody (5 μg) was tested in a native ChIP experiment using chromatin from K-562 cells (3 μg) with the SNAP-ChIP K-AcylStat Panel (EpiCypher Catalog No. 19-3001) spiked-in prior to micrococcal nuclease digestion. Specificity (left Y-axis)was determined by qPCR for the DNA barcodes corresponding to modified nucleosomes in the SNAP-ChIP panel (x-axis). Black bar represents antibody efficiency (right y-axis; log scale) and indicates percentage of the target immunoprecipitated relative to input.
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13-0040 Luminex Data

Luminex Data: Histone H3K4me1 antibody was assessed using a Luminex® based approach employing dCypher™ Nucleosome K-MetStat Panel (EpiCypher Catalog No. 16-9001). The panel comprises biotinylated designer nucleosomes (X-Axis) individually coupled to uniquely identifiable Luminex Magplex® beads. Antibody binding to nucleosomes were tested in multiplex (24-plex) at a 1:1000 dilution, and detected with second layer anti-IgG*PE. Data was generated using a Luminex FlexMAP3D®. Data normalized to relevant on-target (H3K4me1; set to 100) is shown. H
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13-0040 Western Blot data

Western Blot Data: Western blot of acid extracts from HeLa cells untreated ( - ) or treated with sodium butyrate ( + ), using EpiCypher Catalog No. 13-0040 at 0.25 µg/mL.
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13-0040 ICC

Immunocytochemistry Data: Immunocytochemistry of HeLa cells treated with sodium butyrate, using H3K4me1 antibody, EpiCypher Catalog No. 13-0040 (red). Actin filaments have been labeled with fluorescein phalloidin (green).
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