ChIPAb+ NF B p65 (ReIA); clone 12H11

Stock Code: 3586315
Manufacturer Part No: 17-10060

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Descriptions

Analysis Note

ControlIncludes negative control normal mouse IgG and primers specific for human IκBα promoter.

Application

Research Sub CategoryTranscription Factors

This ChIPAb+ NFκB p65 (RelA) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from serum starved, TNFα-treated (20 ng/mL, 30 min) 293 cells (~3 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µg of Normal Mouse IgG or 4 µg of Anti-NFκB p65 (RelA) and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of NFκB p65 (RelA) associated DNA fragments was verified by qPCR using ChIP Primers, IκBα promoter as a positive locus, and β-Actin promoter primers as a negative locus (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Representative lot data.
Huvec lysate (Lane 1), L6 lysate (Lane 2) and PC12 lysate (Lane 3) were resolved by electrophoresis, transferred to PVDF membrane and probed with anti-NFκB p65 (RelA) (1:500 dilution).
Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection
system.
Arrows indicates protein NFκB p65 (RelA) (~65 kDa) (Please see figures).
A non-specific band may be seen at ~230 kDa in L6 lysate.

Immunofluorescence: A 1-10 µg/mL concentration of a previous lot was used in immunofluorescence.

Immunohistochemistry (paraffin sections): A 5-10 µg/mL (APAAP)
concentration of a previous lot was used in immunohistochemistry.

Immunohistochemistry (frozen sections): A 5-10 µg/mL (APAAP)
concentration of a previous lot was used in immunohistochemistry.

Electrophoretic Mobility Shift Assay (EMSA): A 0.5-1 µg/mL concentration of a previous lot was used in shift assay.

Flow Cytometry: A previous lot of this antibody was used in flow cytometry.

Optimal working dilutions must be determined by end user.

Research CategoryEpigenetics & Nuclear Function

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context