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Phospho-NF-kB p65/RelA-S529 Rabbit pAb (AP0944)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of lysates from HeLa cells using Phospho-NF-kB p65/RelA-S529 Rabbit pAb (AP0944) at 1:500 dilution incubated overnight at 4℃. HeLa cells were treated by TNF-α (100 ng/ml) and Calyculin A (50 nM) at 37℃ for 10 minutes.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3 % nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 60s.

Overview

Product name	Phospho-NF-kB p65/RelA-S529 Rabbit pAb
Catalog No.	AP0944
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

NF-kappa-B is a ubiquitous transcription factor involved in several biological processes. It is held in the cytoplasm in an inactive state by specific inhibitors. Upon degradation of the inhibitor, NF-kappa-B moves to the nucleus and activates transcription of specific genes. NF-kappa-B is composed of NFKB1 or NFKB2 bound to either REL, RELA, or RELB. The most abundant form of NF-kappa-B is NFKB1 complexed with the product of this gene, RELA. Four transcript variants encoding different isoforms have been found for this gene.

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S529 of human NF-kB p65/RelA (NP_068810.3).
Sequence	LLSGD
Gene ID
Swiss Prot
Synonyms	p65; CMCU; NFKB3; AIF3BL3; Phospho-NF-kB p65/RelA-S529
Calculated MW	60kDa
Observed MW	65kDa/

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:500 - 1:2000 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 50% glycerol, pH7.3.
Key application	Western blotting
Positive samples	HeLa treated by TNF-α and Calyculin A
Cellular location	Cytoplasm, Nucleus.
Customer validation	WB(Mus musculus, Homo sapiens)