CHD3

CHD3 CUTANA™ CUT&RUN Antibody

$445.00
SKU: 13-2009
Pack size: 100 µL
  • Type:  Polyclonal
  • Host:  Rabbit
  • Target Size:  227 kDa
  • Format:  Aff. Pur. IgG
  • Reactivity:  Human, Mouse
  • Applications: CUT&RUN, IP, IHC

Description

This antibody meets EpiCypher’s “CUTANA Compatible” criteria for performance in Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and/or Cleavage Under Targets and Tagmentation (CUT&Tag) approaches to genomic mapping. Every lot of a CUTANA Compatible antibody is tested in the indicated CUTANA approach using EpiCypher optimized protocols and determined to yield peaks that show a genomic distribution pattern consistent with reported function(s) of the target protein. CHD3 is an understudied member of the nucleosome remodeling and deacetylase (NuRD) complex. CHD3 antibody produces CUT&RUN peaks above background (Figure 1) within gene promoters, intergenic regions, and introns (Figures 1-2).

Validation Data

Figure 1: CHD3 genome-wide enrichment in CUT&RUN
CUT&RUN was performed using 500,000 K562 cells with 0.5 µg CHD3 antibody. (A) Heatmap of CHD3 peaks (MACS2) relative to IgG and H3K4me3 antibodies (0.5 µg; EpiCypher 13-0042 and 13-0041, respectively). Aligned rows are ranked by CHD3 peak intensity (top to bottom) and colored such that red indicates high enrichment and blue denotes background. (B) The number of CHD3 peaks falling within the indicated genomic features is plotted.

Figure 2: Representative CHD3 CUT&RUN peaks
Two gene loci showing CHD3 peaks at a gene promoter (top) and an intronic region (bottom) are representative of the functional annotation analysis of CHD3 peak localization in CUT&RUN (Figure 1). Images were generated using the Integrative Genomics Viewer (Broad Institute).

Figure 3: Immunoprecipitation of human CHD3
EpiCypher CHD3 antibody (3 µg) was used to immunoprecipitate whole cell lysates isolated from HeLa cells using NETN lysis buffer (1.0 mg per IP). A negative control IgG antibody and positive control antibody to a different CHD3 epitope (Bethyl Laboratories) were also used to demonstrate specificity of the IP. Immunoprecipitates were loaded onto 4-8% SDSPAGE gel (20% of IP loaded) and probed via western blot with Bethyl A301-219A antibody (1.0 µg/mL).

Figure 4: Immunohistochemistry detection of human and mouse CHD3
FFPE section of human Ewing sarcoma (left) and mouse renal cell carcinoma (right) examined using CHD3 antibody (1:1,000 dilution, 1 µg/mL).

Technical Information

Immunogen
A synthetic peptide corresponding to human CHD3 amino acids 1950 to 2000.
Formulation
Antigen affinity-purified antibody (1.0 mg/mL) in Tris-citrate/phosphate buffer pH 7 to 8, 0.09% sodium azide.
Storage and Stability
Stable for 1 year at 4°C from date of receipt.

Application Notes

Recommended Dilutions:

CUT&RUN: 0.5 µg

IHC: 1:500 - 1:2,000*

IP: 2 - 5 µg/mg lysate

*Epitope retrieval with citrate buffer pH 6.0 recommended for FFPE tissue

Documents & Resources

Additional Info

This product is provided for commercial sale under license from Bethyl Laboratories, Inc.

Applications Key:

ChIP: Chromatin immunoprecipitation
CUT&RUN: Cleavage Under Targets and Release Using Nuclease
CUT&Tag: Cleavage Under Targets and Tagmentation
E: ELISA
FACS: Flow cytometry
ICC: Immunocytochemistry
IF: Immunofluorescence
IHC: Immunohistochemistry
IP: Immunoprecipitation
L: Luminex
WB: Western Blot

Reactivity Key:

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