CUTANA™

CUTANA™ Rabbit IgG CUT&RUN Negative Control Antibody

$75.00
SKU: 13-0042
Pack Size: 100 µg
  • Type:  Polyclonal
  • Host:  Rabbit
  • Applications:  CUT&RUN
  • Reactivity: Negative Control
  • Format:  Affinity Purified IgG
  • Target Size:  N/A

Description

This Rabbit IgG Antibody can be used to control for non-specific background signal in CUT&RUN and other applications. Pair with the H3K4me3 positive control antibody (EpiCypher 13-0041) for a well-controlled experiment.

Validation Data

Figure 1: CUT&RUN genome wide enrichment
CUT&RUN data was generated using the EpiCypher CUTANA™ CUT&RUN protocol. Sequence reads were aligned to 18,793 annotated transcription start sites (TSSs ± 2 kbp). Signal enrichment was sorted from highest to lowest (top to bottom) relative to H3K4me3 CUT&RUN positive control antibody (EpiCypher 13-0041). High, medium, and low intensity are shown in red, yellow, and blue, respectively. Rabbit IgG negative control antibody displayed minimal background, as expected.

Figure 2: Rabbit IgG CUT&RUN tracks
CUT&RUN data was generated as described in Figure 1. Gene browser shots generated using the Integrative Genomics Viewer (IGV, Broad Institute) show representative loci for H3K4me3 positive control antibody (EpiCypher 13-0041). IgG track displayed minimal background, as expected.

Figure 3: Protein A Immunoprecipitation
To demonstrate functional binding to Protein A, Rabbit IgG Antibody (2 µg) was incubated with Protein A-conjugated magnetic beads in CUT&RUN Antibody Buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM Spermidine, 1x Protease Inhibitor, 0.01% Digitonin, 2 mM EDTA) at 4°C overnight. One-fifth of the unbound flow-through fraction (20% FT) was retained for analysis. The antibody-bead complexes were washed three times with CUT&RUN Digitonin Buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM Spermidine, 1x Protease Inhibitor, 0.01% Digitonin). The antibody was eluted by resuspending beads in 1X SDS Buffer and heating at 95°C for 15min. Antibody was analyzed by SDS-PAGE using Oriole Fluorescent gel staining.

Technical Information

Immunogen
None
Formulation
Affinity-purified antibody in PBS pH 7.6
Storage and Stability
Stable for 1 year at 4°C from date of receipt

Recommended Dilution

Prior to use, prepare a 1 mg/mL working stock dilution in CUT&RUN Antibody Buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM Spermidine, 0.01% Digitonin, 2 mM EDTA). Store working stock at 4°C for 1 month. Use 0.5 µg of the working stock for each CUT&RUN assay. A typical yield for a CUT&RUN assay using 500,000 K562 cells is 2-5 ng DNA.

Documents & Resources

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