Histone

Histone H3K18acyl Antibody, SNAP-ChIP® Certified

$465.00
SKU: 13-0050
Pack Size: 100 µL
  • Type:  Polyclonal
  • Target Size:  15 kDa
  • Format:  Affinity Purified IgG
  • Host:  Rabbit
  • Reactivity:  Human, Mouse, Rat, Yeast
  • Applications:  ChIP, WB, ICC/IF, IHC, IP

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) based on technology originating from Grzybowski et al. [1] and profiling standards from Shah et al. [2]. This antibody reacts to H3K18 acetylation as well as extended acyl states (butyrylation, bu; crotonylation, cr) when present alone and in combination (H3K4,9,14,18ac). No cross reactivity to other lysine acylations in the EpiCypher SNAP-ChIP KAcylStat panel (EpiCypher 19-3001) is detected.

Validation Data

Figure 1: SNAP-ChIP-qPCR
Histone H3K18acyl antibody (3 µg) was tested in a native ChIP experiment using chromatin from K-562 cells (3 µg) with the SNAP-ChIP K-AcylStat Panel 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.

Figure 2: Western Blot Data
Western analysis of H3K18acyl from 30 μg of HeLa whole cell lysate after 0.012% DMSO (Lane A) or 0.4 μM Trichostatin A (Lane B) treatment for 18 hours. H3K18acyl antibody was used for detection at 1:5,000 dilution.

Figure 3: Immunofluorescence
IF detection of histone H3K18acyl in paraformaldehyde fixed HeLa cells. Left Panel: Detection of H3K18acyl (green) using H3K18acyl antibody at a 1:500 dilution and a fluorescently labeled anti-rabbit IgG secondary antibody. Middle Panel: Localization of alpha Tubulin (red). Right Panel: A merged image of the Left and Middle panels with Hoechst dye (blue) indicating nuclear localization of H3K18acyl.

Figure 4: Immunohistochemistry
IHC detection of H3K18acyl in paraffinembedded (A) mouse prostate (B) mouse intestine (C) mouse duodenum and (D) rat forebrain using H3K18acyl antibody at a 1:500 dilution.

Figure 5: Immunoprecipitation
IP analysis of H3K18acyl in HeLa whole cell lysate treated with 0.4 µM Trichostatin A for 18 hours. Lane A: 30 µg of HeLa whole cell lysate. Lane B: IP with control rabbit IgG antibody. Lane C: Protein recovered from IP with H3K18acyl antibody (1:400 dilution). Western analysis was performed by binding with H3K18acyl primary antibody (1:5,000 dilution), followed by detection with an HRP-conjugated anti-rabbit IgG secondary antibody.

Technical Information

Immunogen
A synthetic peptide corresponding to histone H3 acetylated at lysine 18.
Formulation
Antigen affinity-purified polyclonal antibody in PBS pH 7, 1% BSA, 20% glycerol, 0.01% thimerosal.
Storage and Stability
Store at 4°C after thawing. Aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. Stable for 1 year at -20°C from date of receipt.

Recommended Dilution

Chromatin Immunoprecipitation (ChIP): 2 - 5 µg per 1x106 cells

Immunocytochemistry/Immunofluorescence (ICC/IF): 1:100 - 1:1,000

Western Blot (WB): 1:1,000 - 1:10,000

Immunohistochemistry (IHC): 1:100 - 1:1,000

Immunoprecipitation (IP): 1:100 - 1:500

References

Background References:
[1] Grzybowski et al. Mol. Cell (2015). PMID: 26004229
[2] Shah et al. Mol. Cell (2018). PMID: 30244833

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