Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Chromatin Immunoprecipitation [1]
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Validation data
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- Product number
- 44-281G - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-c-Fos (Thr325) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Storage
- -20°C
Submitted references Effects of ATP-competitive and function-selective ERK inhibitors on airway smooth muscle cell proliferation.
Defnet AE, Huang W, Polischak S, Yadav SK, Kane MA, Shapiro P, Deshpande DA
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2019 Oct;33(10):10833-10843
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2019 Oct;33(10):10833-10843
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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.