Anti-DYKDDDDK (FLAG®) tag Rabbit mAb - ChIP, CUT&RUN and CUT&Tag Grade

Overview

Reactivity

All species

Tested applications

WB : 1/1000-1/5000. 

IHC : 1/200-1/500.

ICC/IF : 1/100-1/500.

IP : 1/50-1/200.

ChIP : 1/50-1/100.
CUT&RUN and CUT&Tag
 

Optimal dilutions/concentrations should be determined by the end user.

Product Picture

Specificity

Flag Antibody Recognizes Fusion proteins from all species containing the DYKDDDDK epitope (Binds to same epitope as Sigma's Anti-FLAG M2 Antibody).

Properties

Immunogen

Synthetic peptide: DYKDDDDK conjugated to KLH

Clonality

Monoclonal, clone number:6B2

Isotype

Rabbit IgG

Form

Liquid, 100 μg at 1mg/ml

Storage instruction

store at -20°C 

Applications

WB Image

Lane: E. coli transfected with DYKDDDDK-tagged protein vector at 1:5000.

 

Recommended secondary antibody: Goat anti Rabbit IgG-HRP (Engibody, Catalog number:AT0097)

Recommended ECL: ECL Pico-Detect™ Western Chemiluminescent HRP Substrate (Engibody, Catalog number: IF6747)

ICC/IF Image

Immunocytochemistry/Immunofluorescence: DYKDDDDK Epitope Tag Antibody- Analysis of Chlamydia trachomatis FLAG-tagged IncA (blue) in HeLa cells. Cells were infected with a GFP-expressing strain of Chlamydia trachomatis containing FLAG-IncA under the control of the Tetracycline (Tet) promoter, and either left untreated (0ng/ml Tet) or treated with anhydrotetracycline (10ng/ml Tet). Cells were fixed with 2% paraformaldehyde in PBS for 30 min at RT, permeabilized and blocked in 10% goat serum

 

IP/CoIP Image

Immunoprecipitation:

 DYKDDDDK Epitope Tag Antibody-ChIP Grade - Analysis of 750 ug whole cell lysate/extract of DYKDDDDK-tagged protein expressing 293T cells. 

 

A: 5 ug whole cell lysate/extract of DYKDDDDK-tagged protein expressing 293T cells 

 

B: Control with 2.5 ug of rabbit IgG 

 

C: Immunoprecipitation of DYKDDDDK-tagged protein by 2.5 ug of DYKDDDDK Epitope Tag rabbit monoclonal antibody -ChIP Grade (Cat no: AT1748) 

 

The immunoprecipitated DYKDDDDK-tagged protein was detected by DYKDDDDK Epitope Tag mouse monoclonal antibody (Cat no: AT0022) diluted at 1: 5000. anti-mouse IgG (HRP) was used as a secondary antibody.

ChIP Images

Chromatin Immunoprecipitation: 

Chromatin immunoprecipitations were performed with cross-linked chromatin from 293T cells, untransfected (left) or transfected (right) with a DYKDDDDK-tagged SSRP1 construct and DYKDDDDK Rabbit mAb, SSRP1 Rabbit mAb, or Normal Rabbit IgG, using Enzymatic Chromatin IP Kit (Magnetic Beads). The enriched DNA was quantified by real-time PCR using Human c-Fos Exon 3 Primers and Human EGR1 Intron 3 Primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.