P raf

Manufactured by Santa Cruz Biotechnology

Sourced in United States

P-Raf is a laboratory equipment product manufactured by Santa Cruz Biotechnology. It is a recombinant form of the Raf serine/threonine-protein kinase, which is a key component of the MAPK/ERK signaling pathway.

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Lab products found in correlation

Close spelling variants of this product

P-Raf-1 (6 citations) Anti- p-Raf-1 (2 citations)

4 protocols using p raf

Curcumin-mediated NF-κB Pathway Regulation

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Curcumin and propidium iodide (PI) were purchased from Sigma-Aldrich (Germany). All antibodies (PARP, lamin B, caspase-3, pCREB, catalytic PKA subunits, Ras GAP, pRaf, pErk, NF-κB subunits p50 and p65, RelB, IκBα, actin, and proliferating cell nuclear antigen (PCNA) were obtained from Santa Cruz Biotechnology (USA) and Cell Signaling (USA). Electrophoresis reagents and Bio-Rad protein assay kit were purchased from Bio-Rad Laboratories (USA). The 22-mer double-stranded NF-κB oligonucleotides were obtained from Promega (USA). Dichlorofluorescein diacetate (DCFHDA) was obtained from Molecular Probes (USA). PGE₂, hydroxy PGE₂, and butaprost were purchased from Cayman Chemicals (USA). H89 and PD98059 were obtained from Sigma Aldrich and Cell Signaling. Nitrocellulose membrane and X-ray reagents were purchased from (Amersham Pharmacia Biotech, UK). These chemicals were used according to the manufacturer’s instructions.

Shehzad A., Islam S.U., Lee J, & Lee Y.S. (2014). Prostaglandin E2 Reverses Curcumin-Induced Inhibition of Survival Signal Pathways in Human Colorectal Carcinoma (HCT-15) Cell Lines. Molecules and Cells, 37(12), 899-906.

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Immunoblotting Analysis of Apoptosis Markers

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Whole-cell lysates were prepared with RIPA lysis buffer as described previously [37 (link)]. Approximately 40 mg of protein lysate per sample was loaded and separated in 8–12% SDS-PAGE gel and then transferred to PVDF membrane (Roche, 03010040001); membrane was blocked with 5% non-fat dry milk for 1 h and then incubated with primary antibodies at 4 °C overnight, followed by secondary antibody incubation for 2 h at room temperature and imaging. Primary antibody were purchased from Cell Signaling Technology (CST): Caspase 9 (9508S), Caspase 7 (9492S), Caspase 3 (9662S), PARP (9542 T), Cleaved-Caspase 9 (7237S), Cleaved-Caspase7 (8438S), Cleaved-Caspase 3 (9664S), Cleaved-PARP (5625S), Cytochrome C (Cyt,c, 4280S), Akt (4691S), p-AKT (4060S), Raf 9422S), p-Raf (9421S), Mek (4694S), p-Mek (2338S), Erk (4695S), p-Erk (4377S), Cyclin B1 (4135S), Cdc2 (9116S), p-Cdc2 (4539S), β-actin (4970S), β-actin (MA1–140); secondary antibody goat anti-rabbit IgG-HRP was purchased from Santa Cruz Biotechnology (sc-7074). Western blot bands were quantified using ImageJ with background noise removed using Rolling-Ball correction.

Liu B., Shen Y., Huang H., Croce K.D., Wu M., Fan Y., Liu Y., Xu J, & Yao G. (2020). Curcumin derivative C212 inhibits Hsp90 and eliminates both growing and quiescent leukemia cells in deep dormancy. Cell Communication and Signaling : CCS, 18, 159.

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VEGF-E Homologs Signaling in TNBC

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The TNBC cell line (MDA-MB-231), and human mammary epithelial cells (HMEC) were purchased from ATCC and were maintained in recommended culture condition as per the ATCC instructions^{38 (link)}. The viral VEGF-E homologs (PCPV-VEGF, ORFV-VEGF, and BPSV-VEGF) were a kind gift from Dr. L.M. Wise, University of Otago, Dunedin New Zealand^{39 (link)}. Antibodies against GFP, PCNA, GAPDH, p-Erk, p-MEK, p-Raf, AKT, FoxO1, H2A were purchased from Santa Cruz Biotechnology Inc Dallas, TX, and ERK, PI3k110α, and p-AKT was purchased from Cell Signaling Technologies Inc. Danvers, MA and anti-BrdU antibody from Invitrogen, Thermo Fisher (SI Table 4). The secondary antibodies for western blot are IR conjugated and were from LI-COR Inc. Lincoln, NE, while for microscopy the secondary antibody is Alexa 594 conjugated purchased from Invitrogen Inc. Carlsbad, CA.
LY294002, an inhibitor for PI3K was purchased from Sigma Aldrich Inc. St. Louis, MO. MDA-MB-231 cells were treated with 50 µM and HMEC cells with 30 µM of LY294002 and incubated for 24 h before being used for assays.

Bose D., Banerjee S., Singh R.K., Wise L.M, & Robertson E.S. (2020). Vascular endothelial growth factor encoded by Parapoxviruses can regulate metabolism and survival of triple negative breast cancer cells. Cell Death & Disease, 11(11), 996.

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Western Blot Analysis of Visfatin Signaling

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For Western blotting, JJ012 and SW1353 cells (5 × 10⁵ each) were seeded in six-well plates. After 24 h of treatment with various concentrations of visfatin, the cells were collected and lysed. A total of 50 μg of protein per lane was separated on a 10% SDS-PAGE gel, transferred to a membrane, and probed with primary antibodies, including p-RAF (SC-101791, Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-MEK (2338S, Cell Signaling, Danvers, MA, USA), p-ERK (SC-7383, Santa Cruz Biotechnology, Santa Cruz, CA, USA), HIF-1α (ab1, Abcam, Cambridge, UK), RAF (SC-133, Santa Cruz Biotechnology, Santa Cruz, CA, USA), MEK (SC-6250, Santa Cruz Biotechnology, Santa Cruz, CA, USA), ERK (SC-1647, Santa Cruz Biotechnology, Santa Cruz, CA, USA), VEGF-D (A19242, ABclonal, Woburn, MA, USA), and β-Actin (A5441, Sigma, St. Louis, MO, USA). Appropriate horseradish peroxidase-conjugated secondary antibodies were applied in TBS containing 5% non-fat milk. Protein bands were detected on the blots using enhanced chemiluminescence reagents (GE Healthcare Life Sciences, Glasgow, UK). The data were expressed as a relative expression ratio to β-Actin [37 (link)].

Song C.Y., Hsieh S.L., Yang S.Y., Lin C.Y., Wang S.W., Tsai C.H., Lo Y.S., Fong Y.C, & Tang C.H. (2024). Visfatin Facilitates VEGF-D-Induced Lymphangiogenesis through Activating HIF-1α and Suppressing miR-2277-3p in Human Chondrosarcoma. International Journal of Molecular Sciences, 25(10), 5142.

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