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T-Src is a recombinant protein that represents the Src-family tyrosine kinase, c-Src. It is produced in E. coli and purified for use in biochemical and cell-based assays.

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Lab products found in correlation

P src tyr416, by Cell Signaling Technology (3 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (3 mentions) P src tyr527, by Cell Signaling Technology (2 mentions) Lysis buffer, by Cell Signaling Technology (1 mentions) P pdgfrβ, by Cell Signaling Technology (1 mentions) Pdgf bb, by Merck Group (1 mentions) Vegf a, by Merck Group (1 mentions) P vegfr2, by Abcam (1 mentions) Total vegfr2, by Cell Signaling Technology (1 mentions) Total pdgfrβ, by Cell Signaling Technology (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Gapdh, by Abcam (1 mentions) β actin, by Abcam (1 mentions) Protease inhibitor cocktail, by GenDEPOT (1 mentions) Vinculin, by Merck Group (1 mentions) P tyr clone 4g10, by Merck Group (1 mentions) Sds sample buffer, by Boston BioProducts (1 mentions) Protein a magnetic bead, by Merck Group (1 mentions) Nupage system, by Thermo Fisher Scientific (1 mentions) β actin, by Cell Signaling Technology (1 mentions)

4 protocols using t src

1

Protein Expression and Immunoblotting

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Cells were lysed with lysis buffer (catalog no. #9803s; Cell Signaling Technology) containing protease inhibitor cocktail (catalog no. #3100-001; Gendepot). Total protein was immunoblotted with primary antibodies against KAI1 (catalog no. ab135779; Abcam), LIF (catalog no. AB-449-NA; R&D Systems), tSrc, pSrc (Tyr416), and pSrc (Tyr527) (Cell Signaling Technology), Myc (catalog no. 05-724; Sigma-Aldrich), p53 (catalog no. MABE327; Merck Millipore), Pbx1 (catalog no. sc-889; Santa Cruz), Cav-1 (Caveolin-1, catalog no. sc-894; Santa Cruz), Flot1 (Flotillin1, catalog no. #3253s; Cell signaling Technology), p-VEGFR2 (catalog no. ab38473; Abcam), total VEGFR2 (catalog no. #2479s; Cell Signaling Technology), p-PDGFRβ (catalog no. #3166s; Cell signaling Technology), total PDGFRβ (catalog no. #4564s; Cell signaling Technology), PDGF-BB (catolg no. 07-1437; Sigma-Aldrich), and VEGF-A (catalog no. #ABS82; Merck Millipore) followed by incubation in horseradish peroxidase-conjugated secondary antibodies (Jackson Laboratory). GAPDH (catalog no. ab9485; Abcam) and beta-actin (catalog no. ab8227; Abcam) were used as an internal control.
Lee J.W., Hur J., Kwon Y.W., Chae C.W., Choi J.I., Hwang I., Yun J.Y., Kang J.A., Choi Y.E., Kim Y.H., Lee S.E., Lee C., Jo D.H., Seok H., Cho B.S., Baek S.H, & Kim H.S. (2021). KAI1(CD82) is a key molecule to control angiogenesis and switch angiogenic milieu to quiescent state. Journal of Hematology & Oncology, 14, 148.
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2

Immunoblotting and Immunoprecipitation Protocol

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Cellular lysates
were prepared using RIPA buffer supplemented with protease and phosphatase
inhibitors. The concentration of lysates was determined by Bradford
assay (Bio-Rad). Immunoblotting was performed using the Nupage system
(Life Technologies) with 100 μg of lysate. The primary antibodies
used were DDR2 (Bethyl Laboratories), p-TYR clone 4G10 (Millipore),
t-Src (Cell Signaling Technologies), p-Src family kinase (Tyr416;
Cell Signaling Technologies), β-actin (Cell Signaling Technologies),
and vinculin (Sigma). Immunoprecipitation was performed by incubating
1 mg of lysate with 5 μL of antibody for 1 h at 4 °C with
shaking. Fifty microliters of protein A magnetic beads (Milipore)
was then added, followed by a 2 h incubation at 4 °C with shaking.
The beads were washed three times with RIPA buffer and resuspended
in SDS sample buffer (Boston BioProducts). Immunoblotting was then
performed using the Nupage system (Life Technologies).
Terai H., Tan L., Beauchamp E.M., Hatcher J.M., Liu Q., Meyerson M., Gray N.S, & Hammerman P.S. (2015). Characterization of DDR2 Inhibitors for the Treatment of DDR2 Mutated Nonsmall Cell Lung Cancer. ACS Chemical Biology, 10(12), 2687-2696.
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3

Western Blotting Protein Analysis

Cited 1 time
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Western analysis was performed as described previously (20 (link)). Cell lysates were prepared using complete lysis buffer (EMD Millipore, San Diego, CA) with protease and phosphatase inhibitor cocktails (Roche Diagnostics, Indianapolis, IN). Protein quantification was performed using DC protein assay from Bio-Rad (Hercules, and CA). Western blot analysis was performed as described previously (21 (link),22 (link)). Antibodies used include stromelysin1 (cat. No. 14351-S) dilution 1:1,000 in milk, vascular endothelial cadherin (VE-cadherins; cat No. 2158) dilution 1:1,000 in BSA, P-P38 MAPK (cat No. 9112-S) dilution 1:1,000 in BSA, T-P38MAPK (cat No. 9212-S) dilution 1:1,000 in BSA, P-SRC Tyr-416 (cat No. 6943-S) dilution 1:1,000 in BSA and T-SRC (cat No. 2109-S) dilution 1:1,000 in BSA all from Cell Signaling Technology (Danvers, MA). β-actin (dilution in milk, primary antibodies 1:10,000 and secondary antibodies 1:20,000) from Sigma (St. Louis, MO) and Claudin-5 antibodies (cat No. ab15106) 1:1,000 and secondary antibodies 1:5,000 dilution in milk from Abcam (Cambridge, MA). Band densitometry was done using NIH Image J software.
Kadry R.W., Adil M.S., Newsome A.S, & Somanath P.R. (2021). Cisatracurium attenuates LPS-induced modulation of MMP3 and junctional protein expression in human microvascular endothelial cells. Bioscience trends, 15(1), 50-54.
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4

EL Inhibits Cancer Cell Invasion

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Purified EL (99.2% pure) was purchased from ChromaDex, Inc (Santa Ana, CA). 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-Dephenyltetrazolium Bromide (MTT) was obtained from AbD Serotech (Raleigh, NC). Matrigel® invasion chambers with 8 μm pore size inserts were obtained from BD Biosciences (Bedford, MA). Crystal violet was obtained from Alfa Aesar (Ward Hill, MA). Rhodamine phalloidin was purchased from Thermo Fisher Scientific (Waltham, MA). Antibodies for p-FAKTyr397, t-FAK, p-paxillinTyr118, t-paxillin, p-Src Tyr416, p-Src Tyr527, t-Src, RhoA, p-Rac/Cdc42, t-Rac/Cdc42, GAPDH, and anti-rabbit HRP-conjugated secondary antibody were purchased from Cell Signaling Technology (Danvers, MA). Antibody for vinculin was purchased from EMD Millipore (Billerica, MA). Ki-67 and anti-mouse Alexa 633 fluorophore-conjugated antibodies were purchased from Abcam (Cambridge, MA). DAPI was obtained from Biotium (Fremont, CA). Coverslips were obtained from Carl Zeiss (Ontario, Canada).
Chikara S., Lindsey K., Borowicz P., Christofidou-Solomidou M, & Reindl K.M. (2017). Enterolactone alters FAK-Src signaling and suppresses migration and invasion of lung cancer cell lines. BMC Complementary and Alternative Medicine, 17, 30.
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