Pfra1

Manufactured by Cell Signaling Technology

Sourced in United States, Canada

PFRA1 is a laboratory equipment product offered by Cell Signaling Technology. It serves as a core functional component within the company's product portfolio, designed to facilitate specific research applications. The detailed specifications and intended use of PFRA1 are not available for this factual and unbiased description.

Automatically generated - may contain errors

Lab products found in correlation

9 protocols using pfra1

Immunoblotting Analysis of Cell Signaling

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Samples were prepared as previously stated^{55 (link)}. Horseradish peroxidase-conjugated secondary antibodies (Bio-Rad, Hercules, CA) and enhanced chemiluminescence (ECL) reagent (Amersham Biosciences, Piscataway, NJ) were used for protein detection. Antibodies against phosphorylated-p65, p-MAPK, p-JNK(T183/Y185), p-c-JUN(S73), c-JUN, p-p38, p-Fra-1, Fra-1, SLUG, β-actin, and α-tubulin were purchased from Cell Signaling Technology. Antibodies against E-cadherin and Vimentin were purchased from BD Pharmingen. The antibodies against cytokeratin 18 (CK18) and GAPDH were from Abcam and Advanced Immunochemical Inc, respectively.

Li R., Ong S.L., Tran L.M., Jing Z., Liu B., Park S.J., Huang Z.L., Walser T.C., Heinrich E.L., Lee G., Salehi-Rad R., Crosson W.P., Pagano P.C., Paul M.K., Xu S., Herschman H., Krysan K, & Dubinett S. (2020). Chronic IL-1β-induced inflammation regulates epithelial-to-mesenchymal transition memory phenotypes via epigenetic modifications in non-small cell lung cancer. Scientific Reports, 10, 377.

+ Open protocol

+ Expand

Western Blot Analysis of Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

Whole-cell lysates were prepared in a standard cell extraction buffer (Life Technologies) supplemented with protease and phosphatase inhibitors (Sigma). Lysates were cleared by centrifugation at 20,000 × g for 10 min at 4°C, and total protein concentrations were determined by micro-bicinchoninic assay (Thermo Fisher Scientific). Approximately 20 μg of total protein was loaded per lane on 4%–12% gradient polyacrylamide gels (Thermo Fisher Scientific) under denaturing and reducing conditions and transferred to 0.2 μm nitrocellulose membranes (Bio-Rad). After probing with antibodies, membranes were imaged on a LI-COR Odyssey CLx system. Densitometry measurements were calculated using Image Studio software version 5.2.5 (LI-COR). Antibodies used for western blotting included pEGFR (Tyr 1068, #3777), EGFR (#2232), pMET (Tyr 1234/35, #3126), MET (#8198), pERK (Thr 202/Tyr 204, #4377), ERK (#4695), SPRY2 (#14954), HA (#2367), pFRS2α (Tyr 436, #3861), FGFR1 (#9740), FGFR3 (#4574), p NF-κB (Ser 536, #3033), NF-κB (#8242), pFRA1 (Ser 265, #5841), pAKT (Ser 473, #4060), AKT (#9272), and c-Jun (#9165) from Cell Signaling Technology in addition to GAPDH (sc-32233) from Santa Cruz Biotechnology. Infrared dye-conjugated secondary antibodies for western blotting were purchased from Rockland Immunochemicals.

Day E.K., Sosale N.G., Xiao A., Zhong Q., Purow B, & Lazzara M.J. (2020). Glioblastoma Cell Resistance to EGFR and MET Inhibition Can Be Overcome via Blockade of FGFR-SPRY2 Bypass Signaling. Cell reports, 30(10), 3383-3396.e7.

+ Open protocol

+ Expand

Investigating Anti-Inflammatory Mechanisms

Check if the same lab product or an alternative is used in the 5 most similar protocols

Pd-EE (NIBRGR0000433437) obtained from the Wildlife Natural Products Bank and voucher specimen (KONPVP0000373423) is preserved at the herbarium of the National Institute of Biological Resources (Incheon, Korea). LPS (Escherichia coli 0111:B4), Pam3CSK, poly I:C, polyethyleneimine (PEI), dexamethasone, and ranitidine were purchased from Sigma Chemical Co. (St. Louis, MO, USA). RAW264.7 cells (ATCC number TIB-71) and HEK293 cells (ATCC number CRL-1573) were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA). Fetal bovine serum (FBS), Roswell Park Memorial Institute (RPMI) 1640 medium, Dulbecco’s Modified Eagle’s medium (DMEM), phosphate buffered saline (PBS), TRIzol reagent, and Opti-MEM Reduced Serum Medium were purchased from Gibco (Grand Island, NY, USA). In addition, p38, p-p38, p50, p-p50, p65, p-p65, FRA1, p-FRA1, and Lamin A/C were purchased from Cell Signaling Technology (Beverly, MA, USA), and β-actin was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Kim S.A., Oh J., Choi S.R., Lee C.H., Lee B.H., Lee M.N., Hossain M.A., Kim J.H., Lee S, & Cho J.Y. (2021). Anti-Gastritis and Anti-Lung Injury Effects of Pine Tree Ethanol Extract Targeting Both NF-κB and AP-1 Pathways. Molecules, 26(20), 6275.

+ Open protocol

+ Expand

Evaluation of Alectinib and Brigatinib Synergy

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

APG-2449 was provided by Ascentage Pharm (Rockville, MD). Alectinib and brigatinib were purchased from MedChemExpress (Monmouth Junction, NJ). PS341 was originally provided by Millennium Pharmaceuticals (Cambridge, MA). Human recombinant TRAIL was purchased from PeproTech, Inc. (Rocky Hill, NJ). ERK1/2 and S6 antibodies were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). DR5, p-ERK1/2, Akt, p-Akt, p70S6K, p-p70S6K, 4EBP1, p-4EBP1, p-S6, Fra-1, p-Fra-1, c-Jun, p-c-Jun, c-Fos, p-c-Fos and FosB antibodies were all purchased from Cell Signaling Technology, Inc (Beverly, MA). DR4 antibody (B-N28) was purchased from Cell Sciences (Canton, MA). Other reagents and antibodies were the same as described previously [7] (link).

Zhao W., Yu D., Zhai Y, & Sun S.Y. (2023). ALK inhibitors downregulate the expression of death receptor 4 in ALK-mutant lung cancer cells via facilitating Fra-1 and c-Jun degradation and subsequent AP-1 suppression. Neoplasia (New York, N.Y.), 42, 100908.

+ Open protocol

+ Expand

Immunoblotting of Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein lysates (20 μg/sample) prepared either with RIPA buffer or SDS-PAGE loading buffer were used for immunoblotting with antibodies against FRA1 and actin (Santa CruZ Biotechnology, Santa CruZ, CA), pFRA1, pc-Jun(S73), c-Jun(Ab-243), pERK, pFAK and β1-integrin (Cell Signaling Technology, Danvers, MA), pAKT(Th308) (Genscript, Piscataway, NJ), KIND1 (Sigma) and AKT(Ab-308) (Abcam, Cambridge, MA). The blots were detected with IRDye-conjugated secondary antibodies (Invitrogen), and imaged with the Odyssey Imagining system (Li-COR, Lincoln, NE).

Zhang X., Wu J., Luo S., Lechler T, & Zhang J.Y. (2016). FRA1 promotes squamous cell carcinoma growth and metastasis through distinct AKT and c-Jun dependent mechanisms. Oncotarget, 7(23), 34371-34383.

+ Open protocol

+ Expand

Immunoblotting Analysis of Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoblotting was performed as previously described16 . The following antibodies were used:
pERK1/2 (Thr202/Tyr204, 9106), ERK1/2 (9102), p MEK1/2 (Ser217/221, 9154), MEK1/2
(4694), MEK1 (2352), MEK2 (9147), JUNB (3753), pFRA1 (Ser265, 5841), p-cJUN
(Ser73, 3270) and cJUN (2315) BCL2 (4223) from Cell Signaling Technology;
BRAF^V600E (E19290) from Spring Bioscience; ATK (sc-8312), pAKT
(Ser473, sc-7985-R), FRA1 (sc-28310), BRAF (sc-5284), BCL2 (sc-492) and HSP90
(sc-7947) from Santa Cruz Biotechnology; MITF (ab12039) and AXL (ab89224) from
Abcam; Melan-A (M7196) from Dako; E-Cadherin (610181), Vimentin (550513) and
Fibronectin (610077) from BD Bioscience.

Kong X., Kuilman T., Shahrabi A., Boshuizen J., Kemper K., Song J.Y., Niessen H.W., Rozeman E.A., Geukes Foppen M.H., Blank C.U, & Peeper D.S. (2017). Cancer Drug Addiction is Relayed by an ERK2-Dependent Phenotype Switch. Nature, 550(7675), 270-274.

+ Open protocol

+ Expand

Generation and Characterization of Genetically Engineered Mouse Models

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The generation of p18^−/−, p18^+/−, and p18^−/−:Brca1^+/− mice in Balb/c background, and p18^−/−and p18^−/−;Brca1^f/f; MMTV-Cre (p18^−/−; Brca1^MGKO) mice in Balb/c-B6 mixed background has been previously described [18 (link), 19 (link), 54 (link)]. The Institutional Animal Care and Use Committee at the University of Miami and Shenzhen University approved all animal procedures. Animals were housed in a specific pathogen-free environment with a 12/12 light cycle. Animals were euthanized by exposure to isoflurane followed by cervical dislocation. At least four female mice were analyzed for each genotype, or were transplanted with each type of tumor cells. Histopathology and immunohistochemistry (IHC) were performed as previously described [18 (link), 19 (link), 54 (link)]. The primary antibodies used were: TGFβR1, TGFβR2 (Santa Cruz), p-Smad2, p-FRA1 (Cell signaling), CK5 (Covance), and Vim (Abcam). Immunocomplexes were detected using the Vectastain ABC DAB kit according to the manufacturer’s instructions (Vector Laboratories). The positive results of IHC were quantified by H-score, as previously described [55 (link)].

Bai F., Wang C., Liu X., Hollern D., Liu S., Fan C., Liu C., Ren S., Herschkowitz J.I., Zhu W.G, & Pei X.H. (2022). Loss of function of BRCA1 promotes EMT in mammary tumors through activation of TGFβR2 signaling pathway. Cell Death & Disease, 13(3), 195.

+ Open protocol

+ Expand

Immunoblotting Analysis of Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

Comprehensive Breast Cancer Cell Culture Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

MCF7 and SUM149 cells were cultured per American Type Culture Collection (ATCC) recommendations. Primary murine mammary tumor cells were cultured in phenol-free DMEM/F12 (Gibco), with 10% charcoal-stripped FBS (Gibco), 10 μg/ml insulin, and 10 ng/ml EGF. For treatment of estrogen, AZD5363, and 4OHT, tumor cells were cultured in 10% charcoal-stripped FBS in the presence of E2, AZD5363 (ApexBio Technology), 4OHT (Sigma), or dimethyl sulfoxide (DMSO) for the indicated times and then collected for further analysis. To determine cell viability, 50,000 cells were plated in 24-well plates and treated with DMSO or drugs at the indicated concentrations for 5 days. Viable cell numbers were determined on day 1, day 3, and day 5 by an automatic cell counter (Bio-rad) with trypan blue exclusion. For western blot, tissue and cell lysates were prepared as previously reported [19 (link), 23 (link), 37 (link)]. Primary antibodies used are as follows: HSP90 (Santa Cruz), Gapdh (Ambion), ERα (Santa cruz), Brca1, p-Akt (Ser473), p-4E-bp1 (Thr37/46), p-mTor (Ser2248), p-Gsk3β (Ser9), E-cad, Vim, Snail, Slug, p-Fra1 (Ser 265), p-Rb (ser780) (Cell Signaling) and Fn (Abcam).

Wang C., Bai F., Zhang L.H., Scott A., Li E, & Pei X.H. (2018). Estrogen promotes estrogen receptor negative BRCA1-deficient tumor initiation and progression. Breast Cancer Research : BCR, 20, 74.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!