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Btt 3033

Manufactured by Bio-Techne
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The BTT 3033 is a laboratory instrument designed for the measurement and analysis of various biological and chemical samples. It is a versatile and precise piece of equipment that can be used for a wide range of applications in research and development, quality control, and diagnostic laboratories.

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Lab products found in correlation

Obtustatin, by Bio-Techne (2 mentions) Sulfo nhs lc biotin, by Thermo Fisher Scientific (1 mentions) Puromycin, by GE Healthcare (1 mentions) Ml141, by Bio-Techne (1 mentions) Anti e cadherin ab1416, by Abcam (1 mentions) 3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2h tetrazolium bromide mtt, by Merck Group (1 mentions) Alexa fluor 488 phalloidin, by Cell Signaling Technology (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Bovine serum albumin 4 6 diamidino 2 phenylindole dihydrochloride dapi, by Merck Group (1 mentions) Ps3 synthesizer, by Protein Technologies (1 mentions) Atn 161, by Bio-Techne (1 mentions) Peg400, by Merck Group (1 mentions) Anti phospho akt s473, by Cell Signaling Technology (1 mentions) Anti phospho akt t308, by Cell Signaling Technology (1 mentions) Lentiviral shrna plasmids, by Merck Group (1 mentions) Anti total akt, by Cell Signaling Technology (1 mentions) Anti 4e bp1, by Cell Signaling Technology (1 mentions) Anti gsk3α β, by Cell Signaling Technology (1 mentions) Torin1, by Selleck Chemicals (1 mentions) Anti tsc2, by Cell Signaling Technology (1 mentions)

8 protocols using btt 3033

1

Integrin Signaling Mechanisms in MCF-7 Cells

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To evaluate the integrin signaling mechanism, the proliferation of MCF-7 cells was evaluated in hormone-depleted media with a treatment that consisted of two integrin inhibitors: α2β1, 132 nM BTT 3033 (4724, Tocris), and α1β1, Obtustatin 0.8 nM (4664, Tocris). The cell seeding procedure was the same as described in section 2.6. Culture media were replaced with fresh phenol-free DMEM (supplemented with 1% penicillin–streptomycin and 10% charcoal-stripped FBS ± BTT 3033 or Obtustatin) 24 h after cell seeding. After an incubation period of 72 h, cell proliferation was examined as described in section 2.7.
Reyes-Ramos A.M., Álvarez-García Y.R., Solodin N., Almodovar J., Alarid E.T., Torres-Garcia W, & Domenech M. (2021). Collagen I Fibrous Substrates Modulate the Proliferation and Secretome of Estrogen Receptor-Positive Breast Tumor Cells in a Hormone-Restricted Microenvironment. ACS biomaterials science & engineering, 7(6), 2430-2443.
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2

Integrin Signaling Antibody Protocol

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Anti-α2-integrin (sc-74466), anti-β-Catenin (sc-59737) and anti-RhoGDI (sc-373724) antibodies were from Santa Cruz Biotechnology. Anti-β1-integrin (AB1952), anti-active-β1-integrin (12G10), anti-α3-integrin (MAB2290), anti-p-Src (Y418; 07-909), anti-Src (GD11), anti-GAPDH (MAB374) and anti-GFP (MAB1083) antibodies were from Merck. Anti-p-Shp2 (Y542; 3751) and Shp2 (3752) antibodies were from cell signalling. Anti-E-cadherin (ab1416) and anti-laminin-β1 (ab44941) antibodies were from Abcam. Anti-vinculin (hVIN-1), anti-phosphotyrosine (4G10) and anti-HSC-70 (N69) antibodies were from Sigma-Aldrich. Anti-α4 integrin (MAB1354) was from R&D Systems. Anti-α5-integrin (eBioSAM-1) antibody was from eBioscience. Anti-p-RhoGDI (Y156; OAA100735) antibody was from Aviva Systems Biology. Anti-laminin-α3 (MAB21441) was from Novus Biologicals. Anti-Cdc42 (ACD03) antibody was from cytoskeleton. Anti-mouse horseradish peroxidase (HRP) and anti-rabbit HRP were from Dako. Anti-mouse Alexa 488 and Alexa 568, anti-rabbit Alexa 488 and Alexa 568, Sulfo-NHS-LC-biotin and phalloidin Alexa 647 were all obtained from Thermofisher. BTT-3033, PP2 and ML141 were from Tocris/Biotechne (Bristol, UK); shp099 was from Millipore. α2-integrin and control shRNA vectors were from Sigma-Aldrich. Calyculin A and protease inhibitor cocktail 1 were obtained from Calbiochem. Puromycin was obtained from GE Healthcare.
Howden J.D., Michael M., Hight-Warburton W, & Parsons M. (2021). α2β1 integrins spatially restrict Cdc42 activity to stabilise adherens junctions. BMC Biology, 19, 130.
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3

Integrin α2β1 Inhibitor Assay

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Selective Integrin α2β1 inhibitor: BTT 3033 was from Tocris Bioscience (Bristol, UK). Bovine serum albumin (BSA), 4′,6-Diamidino-2-phenylindole dihydrochloride (DAPI) and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich. All reagents for cell culture were obtained from Invitrogen (Thermo Fisher) except fetal bovine serum (FBS), which was obtained from Millipore. Alexa Fluor 488 Phalloidin was supplied by Cell Signaling.
Rattanasinchai C., Navasumrit P, & Ruchirawat M. (2022). Elevated ITGA2 expression promotes collagen type I-induced clonogenic growth of intrahepatic cholangiocarcinoma. Scientific Reports, 12, 22429.
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4

Inhibitor Screening for hMSC Culture

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hMSCs were exposed to various inhibitor concentrations for 48 h to determine optimal concentrations. The following inhibitors and concentrations were used: 30 μM BTT 3033 (Integrin α2β1 inhibitor, Tocris, Avonmouth, Bristol, U.K.), 30 μM P11 (Integrin αvβ3/αvβ5 inhibitor, Tocris) and 1 mg/mL GAP26 (Connexin-43 peptide-based inhibitor). GAP26 (Val-Cys-Tyr-Asp-Lys-Ser-Phe-Pro-Ile-Ser-His-Val-Arg) peptide was synthesized by standard solid-phase fluorenylmethyloxycarbonyl chloride chemistry on a Rink amide-MBHA resin using PS3 synthesizer (Protein Technologies, Tucson, AZ). These peptides were cleaved and deprotected in trifluoroacetic acid/thioanisole/ethanedithiol/anisole (90/5/3/2). The formation of peptide was characterized by liquid chromatography–mass spectrometry (LC–MS) (Figure S10).
For the inhibition experiment, the indicated concentrations of inhibitors were added to hMSC suspensions before seeding the cells onto the substrates. Cells were cultured for 4 days before harvesting RNA for gene expression analysis.
Balikov D.A., Crowder S.W., Boire T.C., Lee J.B., Gupta M.K., Fenix A.M., Lewis H.N., Ambrose C.M., Short P.A., Kim C.S., Burnette D.T., Reilly M.A., Murthy N.S., Kang M.L., Kim W.S, & Sung H.J. (2017). Tunable Surface Repellency Maintains Stemness and Redox Capacity of Human Mesenchymal Stem Cells. ACS applied materials & interfaces, 9(27), 22994-23006.
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5

Integrin Inhibitors for SARS-CoV-2 Infection

Cited 1 time
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USA-WA1/2020 SARS-CoV-2 strain was obtained from BEI Resources (NIAID, NIH). Integrin inhibitors, BTT3033, a selective inhibitor of α2β1, ATN-161 an integrin α5β1 antagonist27 (link), and GLPG0187 a broad-spectrum integrin inhibitor, were purchased as powders from Tocris Bioscience. The EEA1 rabbit monoclonal antibody (clone C45B10) was from Cell Signaling Technologies (CAT# 3288S). Alexa fluor 647 conjugated F(ab')2 fragment goat anti-rabbit IgG was from Invitrogen (CAT# A21246). In addition, myristoylated peptides; mP6 (Myr-FEEERA-OH) and mP13 (Myr-KFEEERARAKWDT-OH) were custom synthesized at Vivitide.
Simons P., Rinaldi D.A., Bondu V., Kell A.M., Bradfute S., Lidke D.S, & Buranda T. (2021). Integrin activation is an essential component of SARS-CoV-2 infection. Scientific Reports, 11, 20398.
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6

Detailed Signaling Pathway Profiling

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Anti-phospho-S473-Akt, anti-phospho-T308-Akt, anti-Akt1, anti-total Akt, anti-phospho-4E-BP1, anti-4E-BP1, anti-phospho-S6K1, anti-S6K1, anti-phospho-S6, anti-S6, anti-phospho-FoxO3, anti-FoxO3, anti-phospho-GSK3 α/β, anti-GSK3 α/β, anti-phospho-IGF1R, anti-IGF1R, anti-phospho-IRS, anti-IRS, anti-phospho-ERK1/2, anti-ERK1/2, anti-TSC2, anti-mTOR, anti-Rictor, and anti-phospho-tyrosine antibodies were purchased from Cell Signaling Technology. Anti-phospho-S422-SGK and anti-Grb10 antibodies were obtained from Santa Cruz Biotechnology. Rapamycin, Torin1, Ku-0063794, BKM-120, GDC-0941, AZD8931, Erlotinib, BMS754807, OSI906, PF431396, PF573228, and SB273005 were purchased from Selleckchem. Cpd22 and PDGFR inhibitor III were purchased from Calbiochem. Anti-integrin α2, anti-integrin αV, and anti-integrin α5 antibodies for flow cytometry were obtained from BD Biosciences. BTT3033 and anti-integrin α2 antibodies for immunoblot analysis were obtained from Tocris and SantaCruz Biotechnology, respectively. Anti-actin and anti-vinculin antibodies, N-methyl-2-pyrrolidone, PEG400, and lentiviral shRNA plasmids were purchased from Sigma.
Yoon S.O., Shin S., Karreth F.A., Buel G.R., Jedrychowski M.P., Plas D.R., Dedhar S., Gygi S.P., Roux P.P., Dephoure N, & Blenis J. (2017). Focal adhesion- and IGF1R-dependent survival and migratory pathways mediate tumor resistance to mTORC1/2 inhibition. Molecular cell, 67(3), 512-527.e4.
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7

Collagen Gel Assay for Amnion Cell Migration

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Type-1 collagen gel (Cultrex 3-D Culture Matrix Rat Collagen I, #3447-020-01, Trevigen) was prepared according to the manufacture’s protocol. Briefly, collagen I was added and mixed in appropriate volumes of 10X PBS, 1 N NaOH and dH2O in sterile tubes. Blocking antibodies or inhibitors were used as follows: anti-integrin α1 antibody (clone FB12, #MAB1973Z, Millipore), anti-integrin α2 antibody (clone P1E6, #MAB1950Z, Millipore), anti-integrin β1 antibody (clone 6S6, #MAB2253Z, Millipore), Mouse IgG1 control antibody (Clone 107.3, #554721, BD Bioscience), DDR1 inhibitor (DDR1-IN-1, #5077, Tocris), DDR2 inhibitor (LCB 03-0110, #5592, Tocris), integrin α1β1 inhibitor (Obtustatin, #4664, Tocris) and integrin α2β1 inhibitor (BTT 3033, #4724, Tocris). These antibodies or inhibitors were incorporated into gel at indicated concentration. Thereafter, collagen gel was placed to the dishes or plates, where confluent primary amnion cells have been scratched. Dishes or plates were incubated at 37 °C for 20 minutes to promote gel formation. After gel was solidified, medium was gently added with drop-wise in the wells or dishes. Antibodies and inhibitors were also added to the medium at indicated concentration.
Mogami H., Kishore A.H, & Word R.A. (2018). Collagen Type 1 Accelerates Healing of Ruptured Fetal Membranes. Scientific Reports, 8, 696.
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8

Platelet activation agonists and inhibitors

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The following agonists were used: Collagen-related peptide (CRP), with the sequence Gly-Cys-Hyp-(Gly-Pro-Hyp)10-Gly-Cys-Hyp-Gly-NH2, was provided by Dr. Richard W. Farndale, from the University of Cambridge (UK), and crosslinked with SPDP (3-(2-pyridyldithio)propionic acid N-hydroxysuccinimide ester) by Dr. Yotis A. Senis, at the University of Birmingham (UK). Thrombin Receptor Activator Peptide 6 (TRAP-6), with the sequence H-Ser-Phe-Leu-Leu-Arg-Asn-OH, was purchased from Bachem. Collagen Reagent HORM® Suspension (KRH) was purchased from Takeda Austria GmbH (Austria). Thrombin was purchased from Sigma (Sigma-Aldrich, St. Louis, MO).
The following inhibitors were used: Fab fragment of the anti-GPVI monoclonal antibody, OM2, was supplied by Otsuka Pharmaceutical Co., Ltd. (Japan). The strong, selective inhibitor of PAR-1, SCH 79797, was purchased from Santa Cruz Biotechnology (Delaware, CA, USA). The selective inhibitor of integrin α2β1, BTT 3033, was from Tocris Bioscience and was provided by Biogen Científica, S.L. (Spain).
Vélez P., Izquierdo I., Rosa I, & García Á. (2015). A 2D-DIGE-based proteomic analysis reveals differences in the platelet releasate composition when comparing thrombin and collagen stimulations. Scientific Reports, 5, 8198.
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