The protein extracts of lung were incubated with the primary antibody [anti-phosphoinositide 3-kinases (PI3K) (1:1000; Cell Signaling Technology); anti-nuclear factor kappa B (NFκB, p65) (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, U.S.A.); anti-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha (IκBα) (1:10000; Abcam plc); anti-phosphorylated IκBα (1:1000; Cell Signaling Technology); anti-VEGF (1:1000; Santa Cruz Biotechnology); anti-VEGFR-1, -phosphorylated VEGFR-1 (1:1000; Abcam plc); anti-VEGFR-2 (1:500; Millipore Corporation); anti-phosphorylated VEGFR-2 (1:1000; Cell Signaling Technology); anti-Rho-associated kinase (RhoA) (1:1000; Cell Signaling Technology); anti-Akt (1:500, Cell Signaling Technology); anti-phosphorylated Akt (1:2000, Cell Signaling Technology); anti-extracellular signal-regulated kinase (ERK), -phosphorylated ERK (1:3000, Millipore Corporation)]. Then the blots were incubated with the secondary antibody (horseradish peroxidase-conjugated goat anti-mouse IgG antibody, Sigma Chemical Co., St. Louis, MO, U.S.A.). With a computer assisted video densitometer and digitalized software (Kodak Digital ScienceTM ID Image Analysis Software, Eastman Kodak Co., Rochester, NY, U.S.A.), the blots were scanned, photographed then the signal intensity (integral volume) of the appropriate bands were analyzed.
Digital sciencetm id image analysis software
Manufactured by Kodak
Sourced in United States
Kodak Digital ScienceTM ID Image Analysis Software is a software tool designed for analyzing digital images. The core function of this software is to provide users with the ability to perform various image processing tasks, such as measurement, annotation, and data extraction, on digital images.
Automatically generated - may contain errors
Lab products found in correlation
Anti vegfr 2, by Merck Group (2 mentions)
Horseradish peroxidase conjugated goat anti mouse igg antibody, by Merck Group (2 mentions)
Anti vegf, by Santa Cruz Biotechnology (1 mentions)
Anti phosphoinositide 3 kinases pi3k, by Cell Signaling Technology (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti phosphorylated iκbα, by Cell Signaling Technology (1 mentions)
Anti phosphorylated akt, by Cell Signaling Technology (1 mentions)
Anti p enos, by Cell Signaling Technology (1 mentions)
2 protocols using digital sciencetm id image analysis software
1
Measuring Signaling Pathways in Lung Tissue
2
Lung Protein Expression Analysis
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Rats were sacrificed, and their lungs were dissected, frozen in liquid nitrogen immediately, and stored at -80 °C until required. The protein extracts were incubated with the primary antibody [anti-VEGF rabbit polyclonal antibody (Gene Tex, Irvine, CA, USA); anti-VEGFR-2 (1:500; Millipore Corporation, Billerica, MA, USA); anti-p-VEGFR2 rabbit polyclonal antibody (OriGene, Rockville, USA); anti-eNOS, -iNOS (1:1000; Millipore Corporation, Billerica, MA, USA); anti-p-eNOS (1:1000; Cell Signaling Technology, Danvers, MA, USA)]. Then, the blots were incubated with the corresponding secondary antibody (horseradish peroxidase-conjugated goat anti-mouse IgG antibody, Sigma Chemical Co., St. Louis, MO, USA). β-actin expression served as loading control. With a computer-assisted video densitometer and digitalized software (Kodak Digital ScienceTM ID Image Analysis Software, Eastman Kodak Co., Rochester, NY, USA), the blots were scanned, and photographed, then the signal intensity (integral volume) of the appropriate band was analyzed.
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