Glass 8-well Millicel EZslides (Merck Millipore, Ireland) were covered in fibronectin (10 μg/ml, R&D). Fixated cells were blocked and incubated with primary antibodies [αSMA and active ITGA5 (SNAKA51, Millipore, USA)] overnight. Slides were incubated with secondary fluorescent antibodies (Bethyl Laboratories, USA). 4',6-diamidino-2-phenylindole (DAPI) (1000 ng/ml, Abbott, USA) was used for nuclei staining. Actin fibers were stained using Phalloidin-Fluorescein isothiocyanate (FITC) (Tocris Biosciences). Confocal images were taken with a Nikon Ti2E microscope equipped with a Yokogawa W1 spinning disk system and a Plan Apo 60x oil NA1.4 objective using a 405 nm and 561 nm laser. Images were analyzed using Fiji.
Snaka51
Manufactured by Merck Group
Sourced in United States
SNAKA51 is a laboratory instrument designed for scientific research and analysis. It is a versatile piece of equipment that can be used for a variety of applications. The core function of SNAKA51 is to perform advanced data processing and analysis tasks, enabling researchers to extract meaningful insights from their experiments.
Automatically generated - may contain errors
Lab products found in correlation
Fibronectin, by R&D Systems (1 mentions)
Millicel ez slides, by Merck Group (1 mentions)
Phalloidin fluorescein isothiocyanate fitc, by Bio-Techne (1 mentions)
Ti2e microscope, by Yokogawa (1 mentions)
4 6 diamidino 2 phenylindole dapi, by Abbott (1 mentions)
Hb 243, by American Type Culture Collection (1 mentions)
Alexa fluor 488 546 647, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Santa Cruz Biotechnology (1 mentions)
2 protocols using snaka51
1
Immunostaining of Focal Adhesions
2
Integrin-targeting Antibodies for Microscopy
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Function blocking antibodies directed against α5 integrin, clone IIA1 was from BD Biosciences and against αv integrin, clone 69.6.5 was produced and purified as previously described [34] . Following antibodies were used for confocal microscopy. Conformation dependent LIBS anti-α5 antibody clone SNAKA-51 was from Millipore. Phycoerythrin-conjugated anti-α5 antibody (clone SAM-1) was from BioLegend. Anti-β1 antibody clone TS2/16 was produced from the hybridoma cell line TS2/16.2.1 (HB-243, ATCC) and purified on protein-G-sepharose column (Pierce). Anti-αv antibody, clone PLW7, was from Santa Cruz Biotechnology, anti-fibronectin antibody, clone 10, was from BD Biosciences, anti-pFAK-tyr397, clone 141-9 from Life Technologies. Fluorescently labeled secondary antibodies were purchased from Invitrogen (Alexa Fluor -488; -546; -647). Dapi was purchased from Santa Cruz Biotechnology. Human fibronectin stabilized solution was purified as described in Reference 35. Cell culture medium and reagents were from Lonza. All other reagents were of molecular biology quality.
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