Histone

Histone H3K14ac Antibody, SNAP-ChIP® Certified

$465.00
SKU: 13-0049
Pack Size: 100 µL
  • Type:  Recombinant Monoclonal
  • Target Size:  15 kDa
  • Format:  Affinity Purified IgG
  • Host:  Rabbit
  • Reactivity:  Human, Mouse, Rat
  • Applications:  ChIP, WB, IF, IHC

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 H3K14ac alone and within tetraacetyl nucleosomes (H3K4,9,14,18ac) with no cross reactivity to other lysine acylations in the EpiCypher SNAP-ChIP K-AcylStat panel (Catalog No. 19-3001) is detected.

Validation Data

Figure 1: SNAP-ChIP-qPCR
Histone H3K14ac 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 H3K14ac on Jurkat cell lysate (Lane 1), rat liver tissue lysate (Lane 2), and mouse lung tissue lysate (Lane 3) at a 1:500 dilution.

Figure 3: Immunofluorescence
IF detection of H3K14ac in HeLa, MCF7, and PC-3M cells (green) with DAPI as a nuclear counter stain (blue). Anti-H3K14ac was added at a 1:50 dilution, followed by the addition of an anti-rabbit IgG conjugated to a fluorophore.

Figure 4: Immunohistochemistry
IHC detection of H3K14ac in paraffin-embedded human kidney tissue (Left Panel) or rat brain tissue (Right Panel). The sections were pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes, blocked with 5% BSA, and probed using anti- H3K14ac at a 1:50 dilution. Detection was performed with an HRP conjugated compact polymer system, DAB as the chromogen, and the tissue was counter stained with hematoxylin.

Technical Information

Immunogen
A synthetic peptide corresponding to histone H3 acetylated at lysine 14.
Formulation
Protein A affinity-purified recombinant monoclonal antibody in TBS pH 7.4, 0.05% BSA, 40% glycerol, 0.05% sodium azide.
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): 1:50 - 1:1,000

Western Blot (WB): 1:500 - 1:2,000

Immunohistochemistry (IHC): 1:50 - 1:200

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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