BRG1/SMARCA4

BRG1/SMARCA4 CUTANA™ CUT&RUN Antibody

$445.00
SKU: 13-2002
Pack size: 100 µL
  • Type:  Polyclonal
  • Host:  Rabbit
  • Target Size:  185 kDa
  • Format:  Affinity Purified IgG
  • Reactivity:  Human, Mouse
  • Applications: CUT&RUN, WB, 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. BRG1 antibody produces CUT&RUN peaks primarily flanking transcription start sites (TSSs, Figure 1). BRG1 peaks show a large degree of overlap with BRD4 peaks (Figure 2), as has been reported in the literature (Conrad et al., 2017).

Validation Data

Figure 1: FOXA1 peaks in CUT&RUN
CUT&RUN was performed using 500,000 MCF7 cells with 0.5 µg FOXA1 antibody. Peaks were called using MACS2. (A) Heatmap showing FOXA1 peaks relative to IgG negative control antibody (EpiCypher 13-0042) in aligned rows ranked by intensity (top to bottom) and colored such that red indicates high localized enrichment and blue denotes background signal. (B) The number of peaks which fall into distinct classes of functionally annotated genomic regions is plotted.

Figure 2: BRG1 CUT&RUN peak enrichment and functional overlap
The CUT&RUN data from Figure 1 was subjected to peak calling using MACS2. BRG1 peaks overlapped with BRD4 antibody CUT&RUN peaks (EpiCypher 13-2003, top), as has been demonstrated in the literature (Conrad et al., 2017). Three representative loci show BRG1 peaks in relation to control and BRD4 antibodies (bottom, Integrative Genomics Viewer).

Figure 3: Western blot detection of human BRG1.
Whole cell lysates were isolated from HeLa, HEK293T (”293T”), and Jurkat cells using NETN lysis buffer. Lysates (15 µg) were loaded onto 4-8% SDS-PAGE gel and analyzed under standard western blot conditions using BRG1 antibody (0.1 µg/mL).

Figure 4: Western blot detection of mouse BRG1
Whole cell lysates were isolated from NIH 3T3 (“3T3”) cells using NETN lysis buffer. Lysates (15 µg) were loaded onto 4-8% SDS-PAGE gel and analyzed under standard western blot conditions using BRG1 antibody (0.1 µg/mL).

Figure 5: Immunoprecipitation of human BRG1
EpiCypher BRG1 antibody (3 µg) was used to immunoprecipitate whole cell lysates isolated from HeLa cells using NETN lysis buffer (1 mg per IP). A negative control IgG antibody and positive control antibody to a different BRG1/SMARCA4 epitope (Bethyl Laboratories) were also used to demonstrate specificity of the IP. Immunoprecipitates were loaded onto 4-20% SDS-PAGE gel (20% of IP loaded) and probed via western blot with EpiCypher BRG1 antibody (1 µg/mL).

Figure 6: Immunohistochemistry detection of human and mouse BRG1
(A) FFPE section of human ovarian carcinoma examined using BRG1 antibody (1:1,000 dilution, 0.2 µg/mL). (B) FFPE section of mouse renal cell carcinoma examined using BRG1 antibody (1:1,000 dilution, 0.2 µg/mL).

Technical Information

Immunogen
A synthetic peptide corresponding to human BRG1 amino acids 75 to 125.
Formulation
Antigen affinity-purified antibody (200 µg/mL) in Tris-buffered saline with 0.1% BSA and 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

Western Blot (WB): 1:2,000 - 1:10,000

Immunoprecipitation (IP): 2 - 5 µg/mg lysate

Immunohistochemistry (IHC): 1:250 - 1:2000*

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

References

Background References:
Conrad et al. Mol. Cell (2017). PMID: 28844864

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
Current stock: 0