NF-κB p65 (D14E12) XP^® Rabbit mAb #8242

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WB
IP
IHC
IF
F
ChIP
C&R

Supporting Data

REACTIVITY	H M R Hm Mk Dg
SENSITIVITY	Endogenous
MW (kDa)	65
Source/Isotype	Rabbit IgG

Application Key:

WB-Western Blotting
IP-Immunoprecipitation
IHC-Immunohistochemistry
IF-Immunofluorescence
F-Flow Cytometry
ChIP-Chromatin Immunoprecipitation
C&R-CUT & RUN

Species Cross-Reactivity Key:

H-Human
M-Mouse
R-Rat
Hm-Hamster
Mk-Monkey
Dg-Dog

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 10⁶ cells) per IP. This antibody has been validated using SimpleChIP^® Enzymatic Chromatin IP Kits.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

Application	Dilution
Western Blotting	1:1000
Simple Western™	1:10 - 1:50
Immunoprecipitation	1:100
Immunohistochemistry (Paraffin)	1:400 - 1:1600
Immunofluorescence (Immunocytochemistry)	1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized)	1:400 - 1:1600
Chromatin IP	1:100
Chromatin IP-seq	1:100
CUT&RUN	1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #69994.

Protocol

Specificity / Sensitivity

NF-κB p65 (D14E12) XP^® Rabbit mAb recognizes endogenous levels of total NF-κB p65/RelA protein. It does not cross react with other NF-κB/Rel family members. Non-specific staining was observed in pancreatic islets by immunohistochemistry.

Species Reactivity:

Human, Mouse, Rat, Hamster, Monkey, Dog

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu498 of human NF-κB p65/RelA protein.

Background

Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11).

Database Links

UniProt ID: Q04206

Entrez-Gene Id: 5970

限制使用

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For Research Use Only. Not For Use In Diagnostic Procedures.

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NF-κB p65 (D14E12) XP® Rabbit mAb #8242