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 Appl.:  CUT&RUN, WB, IP
Format:  Aff. Pur. IgG Reactivity: Human
BRM/SMARCA2 CUTANA™ CUT&RUN Antibody 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 (EpiCypher.com/resources/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).

BRM/SMARCA2 CUTANA™ CUT&RUN Antibody Immunogen: A synthetic peptide corresponding to human BRM amino acids 1 to 50.

BRM/SMARCA2 CUTANA™ CUT&RUN Antibody Formulation: Antigen affinity-purified antibody (200 µg/mL) in Tris-buffered saline with 0.1% BSA and 0.09% sodium azide.

BRM/SMARCA2 CUTANA™ CUT&RUN Antibody Storage and Stability: Stable for 1 year at 4°C from date of receipt.

BRM/SMARCA2 CUTANA™ CUT&RUN Antibody Application Notes
Recommended Dilutions:
CUT&RUN: 0.1 - 0.5 µg
WB: 1:2,000 - 1:10,000
IP: 2 - 5 µg/mg lysate

BRM/SMARCA2 CUTANA™ CUT&RUN Antibody References
1. Raab et al (2017) Epigenetics Chromatin 10:62.

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

View technical datasheet for this product. 13-2006 Datasheet

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.cerevesiae; Sp-S. pombe; WR-Wide Range (predicted); X-Xenopus; Z-Zebrafish

13-2006 Heatmap

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.
(Click image to enlarge)

13-2006 seq tracks

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).
(Click image to enlarge)

13-2006 WB

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).
(Click image to enlarge)

13-2006 IP

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).

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