BRM/SMARCA2

BRM/SMARCA2 CUTANA™ CUT&RUN Antibody

$445.00
SKU: 13-2006
Pack Size: 100 µL
  • Type:  Polyclonal
  • Host:  Rabbit
  • Target Size:  181 kDa
  • Format:  Affinity Purified IgG
  • Reactivity:  Human
  • Applications: CUT&RUN, WB, IP

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. BRM antibody produces CUT&RUN peaks above background (Figure 1) localized to gene transcription start sites (Figures 1-2), cconsistent with its known role as the ATP-dependent helicase subunit of the SWI/SNF chromatin remodeler complex (1).

Validation Data

Figure 1: BRM enrichment at annotated transcription start sites (TSSs) in CUT&RUN
CUT&RUN was performed using 500,000 K562 cells with BRM (0.1 µg) and control antibodies (0.5 µg; IgG, EpiCypher 13-0042; H3K4me3, EpiCypher 13-0041). Sequencing reads were aligned to TSSs (+/- 2 kbp) of 18,793 genes. Signal (red) over background (blue) is ranked by intensity (top to bottom). All rows aligned to BRM antibody with moderate fixation (0.1% formaldehyde, 1 min), which improved signal vs. native conditions.

Figure 2: BRM CUT&RUN peaks and functional overlap
Two representative gene loci from the CUT&RUN data in Figure 1 are shown. BRM enrichment overlaps with H3K4me3 peaks, consistent with the reported function of BRM as a member of the SWI/SNF chromatin remodeler complex (1). Improved signal recovery with moderate fixation (0.1% formaldehyde, 1 min), is notable. Images generated in Integrative Genomics Viewer (Broad Institute).

Figure 3: Western blot detection of human BRM
Whole cell lysates were isolated from HeLa cells using NETN lysis buffer. The indicated amounts (µg) of lysate were loaded onto 4-8% SDS-PAGE gel and analyzed under standard western blot conditions using BRM antibody (0.04 µg/mL).

Figure 4: Immunoprecipitation of human BRM
EpiCypher BRM 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 antibodies to various BRM epitopes (Bethyl Laboratories) were also used to demonstrate specificity of the IP. Immunoprecipitates were loaded onto 4-8% SDS-PAGE gel (20% of IP loaded) and probed via western blot with EpiCypher BRM antibody (1.0 µg/mL).

Technical Information

Immunogen
A synthetic peptide corresponding to human BRM amino acids 1 to 50.
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: 0.1 - 0.5 µg

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

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

References

Background References:
Raab et al. Epigenetics & Chromatin (2017).

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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