SNF2H/SMARCA5

SNF2H/SMARCA5 CUTANA™ CUT&RUN Antibody

$445.00
SKU: 13-2007
Pack Size: 100 µL
  • Type:  Polyclonal
  • Host:  Rabbit
  • Target Size:  122 kDa
  • Format:  Affinity Purified IgG
  • Reactivity:  Human (predicted: Mouse)
  • 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. SNF2H antibody produces CUT&RUN peaks above background (Figure 1) that overlap with H3K4me3 (Figures 1-2), consistent with its known role as the ATP-dependent helicase subunit of the ISWI chromatin remodeler complex [1].

Validation Data

Figure 1: SNF2H enrichment at annotated transcription start sites (TSSs) in CUT&RUN
CUT&RUN was performed using 500,000 K562 cells with SNF2H (0.1 µg) and control antibodies (0.5 µg; IgG negative control, EpiCypher 13-0042; H3K4me3 positive control, EpiCypher 13-0041). Sequencing reads were aligned to annotated TSSs (+/- 2 kbp) of 18,793 genes. High, medium, and low signal is ranked by intensity (top to bottom) and reflected by red, yellow, and blue colors, respectively. Rows aligned relative to SNF2H antibody.

Figure 2: SNF2H CUT&RUN peaks and functional overlap
Two representative gene loci from the CUT&RUN data in Figure 1 are shown. SNF2H enrichment overlaps with H3K4me3 CUT&RUN peaks (EpiCypher 13-0041), consistent with the reported function of SNF2H as a subunit of ISWI chromatin remodeler complex (1). Images were generated using the Integrative Genomics Viewer (Broad Institute).

Figure 3: Western blot detection of human SNF2H
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 SNF2H antibody (0.04 µg/mL).

Figure 4: Immunoprecipitation of human SNF2H
EpiCypher SNF2H 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 different SNF2H 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 SNF2H antibody (1.0 µg/mL).

Technical Information

Immunogen
A synthetic peptide corresponding to human SNF2H amino acids 50 to 100.
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:
[1] Santos-Rosa et al. Mol. Cell (2003). PMID: 14636589

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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