Histone

Histone H4K5ac Antibody, SNAP-ChIP® Certified

$465.00
SKU: 13-0051
Pack Size: 100 µg
  • Type:  Mixed Monoclonal*
  • Target Size:  11 kDa
  • Format:  Affinity Purified IgG
  • Host:  Rabbit
  • Reactivity:  Human (Predicted: Mouse, Rat, Bovine)
  • Applications:  ChIP, Luminex, ICC/IF

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 H4K5ac when present alone and in combination (H4K5,8,12,16ac). No cross reactivity to other lysine acylations in the EpiCypher SNAP-ChIP K-AcylStat panel (EpiCypher 19-3001) is detected.

*Mixed Monoclonal: a pool of multiple monoclonal antibodies.

Validation Data

Figure 1: SNAP-ChIP-qPCR
Histone H4K5ac 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: Luminex Data
Histone H4K5ac antibody was profiled using a Luminex® based approach utilizing dCypher™ Nucleosome K-AcylStat Panel (Catalog No. 16-9003). The panel comprises biotinylated designer nucleosomes (x-axis) individually coupled to uniquely identifiable Luminex MagPlex® beads. Antibody binding to nucleosomes was tested in multiplex (23-plex) at a 1:250 dilution, and detected with second layer anti-IgG*PE. Data was generated using a Luminex FlexMAP3D®. Data normalized to relevant on-target is shown (H4K5ac; set to 100).

Figure 3: Immunofluorescence
Representative images (60X magnification) of HeLa cells fixed, permeabilized, and immunostained to show endogenous localization of H4K5ac. Panel a: H4K5ac antibody (green, 1:100 dilution) detected with an Alexa Fluor® 488 conjugated anti-rabbit secondary. Panel b: DAPI stained nuclei (blue). Panel c: Rhodamine stained cytoskeletal F-actin (red). Panel d: A composite of panels a, b & c demonstrating the nuclear localization of H4K5ac. Panel e: Negative control lacking H4K5ac primary antibody to assess background.

Technical Information

Immunogen
A synthetic peptide corresponding to histone H4 acetylated at lysine 5 (aa 1 - 9).
Formulation
Protein A affinity-purified recombinant polyclonal antibody in PBS pH 7.4, 0.09% 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

Luminex: 1:250 - 1:4,000

Immunocytochemistry/Immunofluorescence (ICC/IF): 1:100

References

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

Documents & Resources

Current stock: 0