NCOA3/SRC3

NCOA3/SRC3 CUTANA™ CUT&RUN Antibody

$445.00
SKU: 13-2013
Pack size: 100 µL
  • Type:  Polyclonal
  • Host:  Rabbit
  • Target Size:  155 kDa
  • Format:  Aff. Pur. IgG
  • Reactivity:  Human
  • Applications: CUT&RUN, WB

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. Consistent with its role as a nuclear receptor coactivator that facilitates ER-mediated transcription (1), NCOA3 (SRC3) antibody shows CUT&RUN peaks in response to estradiol stimulation (Figure 1) that overlap with known estrogen response element (ERE) binding motifs (Figure 2). SRC3 CUT&RUN peaks also overlap with ER alpha antibodies (N- and C-term; 13-2011 and 13-2012 respectively) (Figure 2).

Validation Data

Figure 1: SRC3 enrichment in CUT&RUN
Serum-starved MCF7 cells were treated with 100 nM estradiol (E2) or vehicle control for 45 minutes. CUT&RUN was performed using 500,000 cells with 0.5 µg of each indicated antibody (gray text). Heatmap shows CUT&RUN enrichment in aligned rows ranked by intensity (top to bottom; relative to ER alpha C-term (13-2012)). Red indicates high localized enrichment and blue denotes background signal.

Figure 2: SRC3 peak analysis in CUT&RUN
Peaks from the E2- treated samples in Figure 1 were called using MACS2. (A) The number of SRC3 peaks which fall into distinct classes of functionally annotated genomic regions is plotted. (B) Homer analysis determined that the ERE consensus motif, represented as a sequence logo position weight matrix, was enriched under SRC3 peaks. (C) The number of SRC3 peaks containing consensus motifs from panel B is shown by Venn Diagram. (D) The number of SRC3 peaks that overlap with ER alpha C-term (13-2012) and ER alpha N-term (13-2011) antibodies are represented by Venn Diagram.

Figure 3: Western blot detection of human SRC3
Whole cell lysates were isolated from HeLa and HEK293T cells using NETN lysis buffer. Lysate (50 µg) was loaded onto 4-8% SDS-PAGE gel and analyzed under standard western blot conditions using SRC3 antibody (0.1 µg/mL).

Technical Information

Immunogen
A synthetic peptide corresponding to human NCOA3/SRC3 amino acids 900 - 950.
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

WB: 1:2000 - 1:10,000

References

Background References:
1. Wagner et al. BMC Cancer (2013). PMID: 24304549

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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