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.
Anti phosphoinositide 3 kinases pi3k
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-phosphoinositide 3-kinases (PI3K) are a family of enzymes responsible for the phosphorylation of phosphoinositides, a crucial step in various cellular signaling pathways. These enzymes play a central role in regulating processes such as cell growth, proliferation, and survival.
Automatically generated - may contain errors
Lab products found in correlation
Anti vegf, by Santa Cruz Biotechnology (2 mentions)
Horseradish peroxidase conjugated goat anti mouse igg antibody, by Merck Group (2 mentions)
Anti vegfr 2, by Merck Group (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)
Digital sciencetm id image analysis software, by Kodak (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Anti protein kinase b akt, by Cell Signaling Technology (1 mentions)
Anti phosphorylated akt p akt, by Cell Signaling Technology (1 mentions)
Anti pten, by Cell Signaling Technology (1 mentions)
Ampkα1, by Abcam (1 mentions)
Anti cox 2, by Abcam (1 mentions)
Digital science id image analysis software, by Kodak (1 mentions)
3 protocols using anti phosphoinositide 3 kinases pi3k
1
Measuring Signaling Pathways in Lung Tissue
2
Western Blot Analysis of Signaling Proteins
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Cell lysates were extracted using radioimmunoprecipitation assays (Beyotime, Shanghai, China), fractionated by electrophoresis, and transferred onto a polyvinylidene fluoride membranes (Millipore). The membranes were blocked with 5% non-fat skim milk and incubated with primary and secondary antibodies. The bands on the membranes were visualized using electrochemiluminescence (Pierce, Rockford, IL, USA). The following primary antibodies were used: anti-KMT2D (1:1,000; Abcam, Cambridge, MA, USA), anti-phosphatase and tensin homolog (PTEN; 1:1,000; Cell Signaling Technology, Danvers, CO, USA), anti-PTEN (1:1,000), anti-phosphoinositide 3-kinases (PI3K) (1:1,000; Cell Signaling Technology), anti-protein kinase B (AKT; 1:1,000; Cell Signaling Technology), and anti-phosphorylated AKT (pAKT; 1:1,000; Cell Signaling Technology).
3
Western Blot Analysis of Protein Markers
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The protein extracts of liver and lung were incubated with the primary antibody [anti-endothelial nitric oxide synthase (eNOS), anti-inducible nitric oxide synthase (iNOS) (1:1000; Millipore Corporation, Billerica, MA, USA); anti-cyclooxygenase (COX)-1 (1:1000; Millipore); anti-COX-2 (1:1000; Abcam plc, Cambridge, UK); anti-5'-adenosine monophosphate-activated protein kinase-α 1 (AMPK-α1) (1:500; ab3759; Abcam plc, Cambridge, UK); anti-VEGF (1:1000; Santa Cruz); anti-VEGF receptor 1 (VEGF-R1) (1:1000; Abcam plc, Cambridge, UK); anti-VEGF receptor 2 (VEGF-R2) (1:500; Millipore Corporation, Billerica, MA, USA); antiphosphoinositide 3-kinases (PI3K) (1:1000; Cell Signaling Technology, Danvers, MA, USA)]. Then the blots were incubated with the secondary antibody (horseradish peroxidase-conjugated goat anti-mouse IgG antibody, Sigma Chemical Co., St. Louis, MO, USA). With a computer assisted video densitometer and digitalizing software (Kodak Digital Science™ 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 bands was analyzed.
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