Log In Sign up
The largest database of trusted experimental protocols

Fgfr3

Manufactured by Cell Signaling Technology
Visit Supplier
Sourced in United States

FGFR3 is a cell surface receptor tyrosine kinase that is a member of the fibroblast growth factor receptor (FGFR) family. It plays a role in the regulation of cell growth, differentiation, and angiogenesis.

Automatically generated - may contain errors

Lab products found in correlation

Fgfr1, by Cell Signaling Technology (6 mentions) Erk1 2, by Cell Signaling Technology (6 mentions) Fgfr4, by Cell Signaling Technology (4 mentions) Bgj398, by Selleck Chemicals (3 mentions) Gefitinib, by Selleck Chemicals (3 mentions) E cadherin, by Cell Signaling Technology (3 mentions) Vimentin, by Cell Signaling Technology (3 mentions) P erk1 2, by Cell Signaling Technology (3 mentions) Cyclin d1, by Santa Cruz Biotechnology (2 mentions) Odyssey clx system, by LI COR (2 mentions) Gapdh, by Santa Cruz Biotechnology (2 mentions) Akt1 2 3, by Santa Cruz Biotechnology (2 mentions) Phospho egfr y1068, by Cell Signaling Technology (2 mentions) Phospho akt s473, by Cell Signaling Technology (2 mentions) Azd9291, by Selleck Chemicals (2 mentions) Wz4002, by Selleck Chemicals (2 mentions) Phospho erk1 2t202 y204, by Cell Signaling Technology (2 mentions) Phospho egfr y1068, by Abcam (2 mentions) Phospho frs2αy436, by Cell Signaling Technology (2 mentions) N cadherin, by Cell Signaling Technology (2 mentions)

Close spelling variants of this product

Anti-FGFR3 (2 citations)

13 protocols using fgfr3

1

Investigating Oncogenic Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols
Gefitinib (ZD1839), osimertinib (AZD9291), BGJ398, dactolisib (BEZ235) and PD0332991 were purchased from Selleck Chemicals (Houston, TX, USA). Reagents were formulated and stored according to the manufacturer’s protocols. The following primary antibodies were used: AKT (#9272), p-AKT S473 (#9271), mTOR (#2983), p-mTOR S2448 (#2971), S6K (#9205S), p-S6K Thr389 (#9202S), S6 (#2217), p-S6 Ser235/236 (#4858), and FGFR3 (#4574) from Cell Signaling Technology (Danvers, MA, USA); Shisa3 (#167069, for Western blot), FGFR1 (#ab10646) and p-FGFR3 Y724 (#ab155960) from Abcam (Cambridge, MA); p-FGFR1 T653/T654 (#06–1433) from Merck Millipore (Darmstadt, Germany), cyclin D1 (#sc-8396), CDK4 (#sc-23,896), CDK6 (#sc-7961), and p16 (#sc-166,760) from Santa Cruz Biotechnology (Santa Cruz, CA); β-actin (#HRP-60008) from Proteintech Group (Rosemont, IL, USA). The secondary HRP-conjugated goat anti-rabbit (#CW0103S) and anti-mouse antibodies (#CW0102S) were from CWBIO (Beijing, China).
Si J., Ma Y., Bi J.W., Xiong Y., Lv C., Li S., Wu N, & Yang Y. (2019). Shisa3 brakes resistance to EGFR-TKIs in lung adenocarcinoma by suppressing cancer stem cell properties. Journal of Experimental & Clinical Cancer Research : CR, 38, 481.
+ Open protocol
+ Expand
2

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), pNF-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
3

Investigating EGFR and FGF Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following antibodies were used: phospho EGFR Y1068 (Abcam AB5644),
phospho EGFR Y1068 (Cell Signaling 3777), EGFR (Cell Signaling 2646), EGFR (Cell
Signaling 4267), phospho ERK1/2 T202/Y204 (Cell Signaling 9101), phospho ERK1/2,
T202/Y204 (Cell Signaling 4370), ERK1/2 (Cell Signaling 9102), phospho AKT S473
(Cell Signaling 4060), AKT1/2/3 (Santa Cruz sc-8312), BIM (Cell Signaling 2933),
Actin (Cell Signaling 4970), Actin-HRP conjugated (Cell Signaling 12262), FGFR1
(Cell Signaling 9740), and FGFR3 (Cell Signaling 4574), phospho FRS2α
Y436 (Cell Signaling 3861), E-Cadherin (Cell Signaling 3195), N-Cadherin (Cell
Signaling 13116), Zeb1 (Cell Signaling 3396), Vimentin (Cell Signaling 5741).
Gefitinib, WZ4002, AZD9291, and BGJ398 (all from Selleck) were dissolved in DMSO
to a final concentration of 10 mmol/liter and stored at −20 °C.
The 18 drugs tested in the long-term assay are listed in Supplemental Table 3.
Raoof S., Mulford I.J., Frisco-Cabanos H., Nangia V., Timonina D., Labrot E., Hafeez N., Bilton S.J., Drier Y., Ji F., Greenberg M., Williams A., Kattermann K., Damon L., Sovath S., Rakiec D.P., Korn J.M., Ruddy D.A., Benes C.H., Hammerman P.S., Piotrowska Z., Sequist L.V., Niederst M.J., Barretina J., Engelman J.A, & Hata A.N. (2019). Targeting FGFR overcomes EMT-mediated resistance in EGFR mutant non-small cell lung cancer. Oncogene, 38(37), 6399-6413.
+ Open protocol
+ Expand
4

Immunoblotting of FGF Receptor Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total protein lysates were extracted from untreated cell lines as well as 48 h after siRNA transfection or 2 or 5 h after inhibitor treatment. For western blot analysis equal amounts of total protein were loaded on NuPAGE® Bis-Tris Gels (Life Technologies). Electrophoretically separated proteins were transferred to nitrocellulose membrane and unspecific binding was blocked with 5% milk/PBS-Tween. Membranes were incubated with primary antibodies overnight at 4°C (FGFR2 1:500; ACTB 1:5000, AC-15, both Sigma Aldrich; FGFR1 1:500; FGFR4 1:500, AM11076PU-N, both from Acris Antibodies, San Diego CA, U.S.A.; FGFR3 1:500, D2G7E; phospho-FGFR 1:500, 55H2; ERK1/2 1:1000; phospho-ERK1/2 1:1000, 20G11, all from CellSignaling Technology, Danvers MA, U.S.A.; HPRT 1:1000, abcam). After incubation with HRP-conjugated secondary antibody SuperSignal West Pico or Femto Chemiluminescent Substrate (Thermo Scientific) was used for detection.
Künstlinger H., Fassunke J., Schildhaus H.U., Brors B., Heydt C., Ihle M.A., Mechtersheimer G., Wardelmann E., Büttner R, & Merkelbach-Bruse S. (2015). FGFR2 is overexpressed in myxoid liposarcoma and inhibition of FGFR signaling impairs tumor growth in vitro. Oncotarget, 6(24), 20215-20230.
+ Open protocol
+ Expand
5

Antibody Immunoblotting for Cell Cycle and Apoptosis

Check if the same lab product or an alternative is used in the 5 most similar protocols
PTX and BGJ398 were purchased from Selleckchem (Houston, TX, USA). The following antibodies against the following proteins were purchased from Santa Cruz Biotechnology (Dallas, TX, USA): cyclin D1 (sc-753), cyclin B1 (sc-752), cyclin A (sc-751), p27 (sc-1641), PARP (sc-8007), β-actin (sc-130656). Antibodies against Snail (cs#3879), E-cadherin (cs#3195), vimentin (cs#5741), FGFR1 (cs#9740), FGFR3 (cs#4574), p21 (cs#2947), caspase-3 (cs#9668), γ-H2AX (cs#2577), β-actin (cs#4970), and cleaved caspase-9 (cs#7237) were purchased from Cell Signaling Technology (Danvers, MA, USA). rhFGF and rhEGF proteins were purchased from R&D Systems (Minneapolis, MN, USA). All chemicals and reagents used were of analytical and were obtained from commercial sources.
Kim S.H., Ryu H., Ock C.Y., Suh K.J., Lee J.Y., Kim J.W., Lee J.O., Kim J.W., Kim Y.J., Lee K.W., Bang S.M., Kim J.H., Lee J.S., Ahn J.B., Kim K.J, & Rha S.Y. (2018). BGJ398, A Pan-FGFR Inhibitor, Overcomes Paclitaxel Resistance in Urothelial Carcinoma with FGFR1 Overexpression. International Journal of Molecular Sciences, 19(10), 3164.
+ Open protocol
+ Expand
6

Western Blot Analysis of Apoptosis Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysates were prepared using lysis buffer (1% NP-40, 0.1% SDS, 1 mM EDTA, 150 mM NaCl, 50 mM Tris pH7.5, supplemented with Complete Protease Inhibitors (Roche) and PhosSTOP (Roche)). Protein concentration was determined with Bio-Rad Protein Assay Dye (5000006), and lysates were separated by SDS-PAGE followed by blotting onto nitrocellulose membranes and incubation with primary antibody (1:1000 in 5% milk) overnight. After washing in TBS/T and incubation with secondary antibody (Li-Cor IRDye 800CW donkey anti-rabbit, 926-32213, dilution 1:20000), blots were developed on a Li-Cor Odyssey CLx system and acquired using Imagestudio (Li-Cor). The following primary antibodies were used: Actin (A4700, Sigma), BAK (12105, Cell Signaling), BAX (2772, Cell Signaling), BCL-2 (4223, Cell Signaling), ERK1/2 (4695, Cell Signaling), basic FGF (20102, Cell Signaling), FGFR1 (9740, Cell Signaling), FGFR3 (4574, Cell Signaling), FGFR4 (8562, Cell Signaling), GFP (In house), HSP60 (4870, Cell Signaling), MCL-1 (5453, Cell Signaling), pERK1/2 (4370, Cell Signaling), Caspase 3 (9662, Cell Signaling), Caspase 9 (9502, Cell Signaling), PARP1 (9532, Cell Signaling) alpha-Tubulin (T5168, Sigma) and cleaved Caspase 3 (9664, Cell Signaling).
Bock F.J., Sedov E., Koren E., Koessinger A.L., Cloix C., Zerbst D., Athineos D., Anand J., Campbell K.J., Blyth K., Fuchs Y, & Tait S.W. (2021). Apoptotic stress-induced FGF signalling promotes non-cell autonomous resistance to cell death. Nature Communications, 12, 6572.
+ Open protocol
+ Expand
7

Inhibition of HSP90 in B-cell Malignancies

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
A high-affinity HSP90-inhibitor, AUY922 [30 (link)] was purchased from Selleckchem. Antibodies to HSP90αβ, PLCγ2, BCAP, CD19, AXL, BCL2, GAPDH and actin were purchased from Santa Cruz Biotechnologies. Antibodies to CD79a, CD79b, LYN, SYK, BTK, AKT, P-ERK1/2, ERK1/2, STAT3, PTPN22, FGFR3, and MCL-1 were purchased from Cell Signaling Technologies. XIAP antibody, chromogen-conjugated antibodies to human CD5 and CD19 or fluorescein isothiocyanate (FITC)-conjugated annexin V were obtained from BD Biosciences or Invitrogen, respectively. Propidium iodide (PI) and other chemicals were purchased from Sigma or Bio-Rad. Replication-deficient lentiviral constructs expressing HSP90-specific shRNA or GFP tagged control scrambled shRNA were purchased from Santa Cruz Biotechnologies.
Mahmud H., Mendez M., Mukhopadhyay B., Holter-Chakrabarty J, & Ghosh A.K. (2020). HSP90 overexpression potentiates the B-cell receptor and fibroblast growth factor receptor survival signals in chronic lymphocytic leukemia cells. Oncotarget, 11(22), 2037-2046.
+ Open protocol
+ Expand
8

Protein Extraction and Analysis of RMS Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total cellular protein from human RMS cell lines was obtained by lysis in 2% SDS buffer supplemented with protease inhibitors (Santa Cruz Biotechnology). Samples were boiled, vortexed and homogenized by passage through a syringe. 20-50ug of protein was loaded onto 4%–20% Mini-Protean TGX gels (Biorad) and transferred onto PVDF membranes. GTP-bound RHOA and RAC1 were precipitated using pull-down activation assay Biochem Kits, according to manufacturer’s recommendations (Cytoskeleton). Antibodies against VANGL2 (2G4, Dr. Jean-Paul Borg and Millipore), PAX7 (DSHB), MYOG (Santa Cruz, M-225), FGFR3 (Cell Signaling, C51F2), CD133 (Cell Signaling, D2V8Q), GAPDH (Cell Signaling, 14C10), RHOA (Cell Signaling, 67B9), RAC1 (Cytoskeleton, C7H2) were used.
Uninfected or stably-infected cells were plated in flat, clear bottom 96 well plates and fixed at the specified time point in 4% PFA/PBS for 15 minutes at room temperature. Cells were washed in 1x PBS, permeabilized in 0.5% Triton X-100/PBS, washed in 0.1% PBST and incubated with primary antibody overnight. Fluorescently-conjugated secondary antibody was used at 1:1000 dilution with 1:5000 DAPI to stain nuclei. Cells were washed in 1x PBS and imaged using a LSM710 Zeiss Laser scanning confocal microscope. Images were processed using ZEN software and and quantifications were made using ImageJ software.
Hayes M.N., McCarthy K., Jin A., Oliveira M.L., Iyer S., Garcia S.P., Sindiri S., Gryder B., Motala Z., Petur Nielsen G., Borg J.P., van de Rijn M., Malkin D., Khan J., Ignatius M.S, & Langenau D.M. (2018). Vangl2/RhoA Signaling Pathway Regulates Stem Cell Self-Renewal Programs and Growth in Rhabdomyosarcoma. Cell stem cell, 22(3), 414-427.e6.
+ Open protocol
+ Expand
9

Investigating EGFR and FGF Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following antibodies were used: phospho EGFR Y1068 (Abcam AB5644),
phospho EGFR Y1068 (Cell Signaling 3777), EGFR (Cell Signaling 2646), EGFR (Cell
Signaling 4267), phospho ERK1/2 T202/Y204 (Cell Signaling 9101), phospho ERK1/2,
T202/Y204 (Cell Signaling 4370), ERK1/2 (Cell Signaling 9102), phospho AKT S473
(Cell Signaling 4060), AKT1/2/3 (Santa Cruz sc-8312), BIM (Cell Signaling 2933),
Actin (Cell Signaling 4970), Actin-HRP conjugated (Cell Signaling 12262), FGFR1
(Cell Signaling 9740), and FGFR3 (Cell Signaling 4574), phospho FRS2α
Y436 (Cell Signaling 3861), E-Cadherin (Cell Signaling 3195), N-Cadherin (Cell
Signaling 13116), Zeb1 (Cell Signaling 3396), Vimentin (Cell Signaling 5741).
Gefitinib, WZ4002, AZD9291, and BGJ398 (all from Selleck) were dissolved in DMSO
to a final concentration of 10 mmol/liter and stored at −20 °C.
The 18 drugs tested in the long-term assay are listed in Supplemental Table 3.
Raoof S., Mulford I.J., Frisco-Cabanos H., Nangia V., Timonina D., Labrot E., Hafeez N., Bilton S.J., Drier Y., Ji F., Greenberg M., Williams A., Kattermann K., Damon L., Sovath S., Rakiec D.P., Korn J.M., Ruddy D.A., Benes C.H., Hammerman P.S., Piotrowska Z., Sequist L.V., Niederst M.J., Barretina J., Engelman J.A, & Hata A.N. (2019). Targeting FGFR overcomes EMT-mediated resistance in EGFR mutant non-small cell lung cancer. Oncogene, 38(37), 6399-6413.
+ Open protocol
+ Expand
10

Antibody Panel for Cancer Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies against AKT (#9272), FGFR-3 (#4574), FGFR-4 (#8562), p70S6K (#9202), Survivin (#2803), Cdk2 (#2546), Cdc25C (#4688), Cyclin B1 (#4138), cleaved caspase 3 (#9661), cleaved PARP (#5625), Cdc2 (#9112), Rb (#9313), S6R (#2217), 4EBP1 (#9452) and α-tubulin (#2144) were obtained from Cell Signaling Technology, Beverly, MA, USA. Phosphorylation-specific antibodies against AKT Ser473 (#9271), FRS2-α Tyr439 (#3861), p70S6K Thr421/424 (#9204), S6R Ser235./236 (#2211), 4EBP1 Thr70 (#9455), Histone 3 Ser10 (#9701), Cdc2 Tyr15 (#9111), p-mTOR (#2971), p-Rb (#9308) and ERK1/2 Thr202/Tyr204 (#4370) were also obtained from Cell Signaling Technology, Beverly, MA, USA. The antibodies against FGFR-2 (sc-122), ERK1/2 (sc-94), FRS2-α (sc-17841), E2F1 (sc-251) and p27 (sc-528) were obtained from Santa Cruz Biotechnology Inc, Santa Cruz, CA, USA. The antibody LYVE-1 (AB-2988) was obtained from EMD Millipore, Billerica, MA, USA. The anti-mouse CD31 (#2502) antibody was obtained from BioLegend, San Diego, CA, USA. Infigratinib and Sorafenib were purchased from Selleck Chemicals, Houston, TX, USA. Bevacizumab (Avastin) was from Genentech, Inc., South San Francisco, CA, USA.
Le T.B., Vu T.C., Ho R.Z., Prawira A., Wang L., Goh B.C, & Huynh H. (2020). Bevacizumab Augments the Antitumor Efficacy of Infigratinib in Hepatocellular Carcinoma. International Journal of Molecular Sciences, 21(24), 9405.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!