Invitrogen

Phospho-c-Fos (Thr325) Polyclonal Antibody

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This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

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Phospho-c-Fos (Thr325) Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Phospho-c-Fos pThr325 was done on 70% confluent log phase HeLa cell treated with 200 nM of PMA for 20 minutes. The cell were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-c-Fos pThr325 Rabbit Polyclonal Antibody (Product # 44-281G) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear and cytoplasmic localization. Panel e is untreated cell with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Phospho-c-Fos (Thr325) Antibody in Western Blot (WB) — Lysates prepared from A431 cells left untreated (1) or treated with EGF (lanes 2 & 3) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes incubated with c-Fos [pT325] antibody (Product # 44-281G).

Phospho-c-Fos (Thr325) Antibody in ChIP assay (ChIP) — Enrichment of endogenous c-Fos (pT325) Protein at specific gene loci using Anti-c-Fos (pT325) Rabbit Polyclonal Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-c-Fos (pT325) Rabbit Polyclonal Antibody (Product # 44-281G, 3 µg) on sheared chromatin from 2 million A431 cells treated with EGF (200 ng/mL), for 30 minutes using the "MAGnify ChIP system" kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by 7500 Fast qPCR system (Product # 4351106) with optimized PCR primer pairs for the promoter of active IL-6, CDKN1A gene, used as positive control target, and the SAT2, used as negative control target. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Phospho-c-Fos (Thr325) Antibody — Antibody specificity was demonstrated by detection of enrichment of the target protein at specific gene loci. Chromatin Immunoprecipitation (ChIP) was performed using Phospho-c-Fos (Thr325) Rabbit Polyclonal Antibody (Product # 44-281G) with relevant positive (CDKN1A and IL-6) and negative (SAT2) sites. {RE}

Product Details

44-281G

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	-
Immunocytochemistry (ICC/IF)	1:250	-
ChIP assay (ChIP)	1-3 µg x 10^6 cells	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human c-Fos that contains threonine 325. The sequence is conserved in human, mouse and rat. View antigen
Conjugate	Unconjugated
Form	Liquid
Purification	Antigen affinity chromatography
Storage buffer	Dulbecco's PBS, pH 7.3, with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20°C
RRID	AB_2533622

Target Information

FBJ murine osteosarcoma viral oncogene homolog is commonly known as Cellular protooncogene Fos (c-Fos). It belongs to the fos transcription factor family and is encoded by the C-FOS gene located on chromosome 14. c-Fos heterodimerizes with c-Jun to form the Activator Protein -1 (AP-1) transcription factor which regulates transcription of several genes that play an important role in cellular signal transduction, cell proliferation and differentiation. c-Fos has been shown to be phosphorylated by various stimuli including growth factors and insulin at atleast seven different sites. The best known upstream kinase for phosphorylation at Threonine 232 is ERK MAPK which regulates localization of c-Fos to the nucleus and is thus important for c-Fos induced transcriptional activity.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: activator protein 1; cellular oncogene c-fos; Cellular oncogene fos; FBJ murine osteosarcoma viral (v-fos) oncogene homolog (oncogene FOS); FBJ murine osteosarcoma viral oncogene homolog; Fos proto-oncogene, AP-1 trancription factor subunit; G0/G1 switch regulatory protein 7; Proto-oncogene c-Fos

Gene Aliases: AP-1; C-FOS; FOS; G0S7; p55

UniProt ID: (Human) P01100

Entrez Gene ID: (Human) 2353

Molecular Function: basic leucine zipper transcription factor