BAX samples were electrophoretically separated on 4–12% NuPage (Invitrogen) gels, transferred to mobilon-FL PVDF membranes (Millipore) and subjected to immunoblotting. For visualization of proteins with Odyssey Infrared Imaging System (LI-COR Biosciences) membranes were blocked in PBS containing 2.5% milk powder. Primary BAX yth-2d2 antibody (R&D systems) was incubated overnight at 4°C in a 1:1,000 dilution. After washing, membranes were incubated with an IRdye800-conjugated goat anti-mouse IgG secondary antibody (LI-COR Biosciences) in a 1:5,000 dilution. Protein was detected with Odyssey Infrared Imaging System. Densitometry of protein bands were acquired using a LI-COR Odyssey® scanner. Quantification and analysis was performed using the Western Analysis tool from the Image Studio 3.1 software.
Mobilon fl pvdf membranes
Manufactured by Merck Group
Mobilon-FL PVDF membranes are a type of laboratory equipment used for various applications. These membranes are made of polyvinylidene fluoride (PVDF) material and are designed for filtration and separation purposes in research and analytical settings. The core function of Mobilon-FL PVDF membranes is to facilitate the efficient separation and isolation of molecules, proteins, or other analytes from complex samples.
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Lab products found in correlation
Odyssey infrared imaging system, by LI COR (5 mentions)
Nupage, by Thermo Fisher Scientific (5 mentions)
Irdye800 conjugated goat anti mouse igg secondary antibody, by LI COR (3 mentions)
Odyssey scanner, by LI COR (3 mentions)
Primary bax yth 2d2 antibody, by R&D Systems (2 mentions)
Irdye 800 conjugated goat anti rabbit igg, by LI COR (2 mentions)
Mcl 1, by Cell Signaling Technology (2 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (2 mentions)
β actin, by Merck Group (2 mentions)
Cytochrome c, by BD (1 mentions)
Vdac1 porin, by Abcam (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Bcl xl, by BD (1 mentions)
Cox 4, by Cell Signaling Technology (1 mentions)
Irdye 800 conjugated goat anti mouse igg, by LI COR (1 mentions)
Irdye 680rd conjugated goat anti rabbit igg, by LI COR (1 mentions)
Odyssey blocking buffer, by LI COR (1 mentions)
β tubulin, by Cell Signaling Technology (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Bcl xl, by Cell Signaling Technology (1 mentions)
5 protocols using mobilon fl pvdf membranes
1
Quantitative Western Blot Analysis
2
Quantitative Western Blot Analysis of BAX
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BAX samples were electrophoretically separated on 4–12% NuPage (Invitrogen) gels, transferred to mobilon-FL PVDF membranes (Millipore), and subjected to immunoblotting.(For visualization of proteins with Odyssey Infrared Imaging System (LI-COR Biosciences), membranes were blocked in PBS containing 2.5% milk powder. Primary BAX monoclonal antibody (Cell Signaling, Cat. #2772) was incubated overnight at 4 °C in a 1:1000 dilution. After washing, membranes were incubated with an IRdye800-conjugated goat anti-mouse IgG secondary antibody (LI-COR Biosciences, cat. # 925-32210) in a 1:5000 dilution. Protein was detected with Odyssey Infrared Imaging System. Densitometry of protein bands was acquired using an LI-COR Odyssey scanner. Quantification and analysis were performed using the western analysis tool from the Image Studio 3.1 software.
3
Immunoblotting Analysis of Apoptosis-Related Proteins
4
Quantitative Western Blot Analysis
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BAX samples were electrophoretically separated on 4–12% NuPage (Invitrogen) gels, transferred to mobilon-FL PVDF membranes (Millipore) and subjected to immunoblotting. For visualization of proteins with Odyssey Infrared Imaging System (LI-COR Biosciences) membranes were blocked in PBS containing 2.5% milk powder. Primary BAX yth-2d2 antibody (R&D systems) was incubated overnight at 4°C in a 1:1,000 dilution. After washing, membranes were incubated with an IRdye800-conjugated goat anti-mouse IgG secondary antibody (LI-COR Biosciences) in a 1:5,000 dilution. Protein was detected with Odyssey Infrared Imaging System. Densitometry of protein bands were acquired using a LI-COR Odyssey® scanner. Quantification and analysis was performed using the Western Analysis tool from the Image Studio 3.1 software.
5
Western Blot Analysis of Apoptosis Regulators
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Protein lysates were obtained by cell lysis in 1% NP-40 buffer (50 mM Tris-HCL, 150 mM NaCl, 1 mM EDTA, 10% Glycerol, 1% NP-40, pH 7.50). Protein samples were electrophoretically separated on 4–12% NuPage (Life Technologies) gels, transferred to mobilon-FL PVDF membranes (Millipore), and subjected to immunoblotting. For visualization of proteins with Odyssey Infrared Imaging System (LI-COR Biosciences) membranes were blocked in Odyssey Blocking Buffer (LI-COR Biosciences). Primary antibodies were incubated overnight at 4 °C in a 1:1,000 dilution. After washing, membranes were incubated with an IRDye800-conjugated goat anti-rabbit IgG (1:10,000 dilution) or IRDye800-conjugated goat anti-mouse IgG (1:20,000 dilution) or IRDye680RD-conjugated goat anti-Rabbit IgG (1:20,000 dilution) secondary antibodies (LI-COR Biosciences). Proteins were detected with Odyssey Infrared Imaging System. Antibodies were used to detect the following proteins on membrane: BCL-XL (Cell Signaling Cat. 2762), MCL-1 (Cell Signaling Cat. 4572), BAX (Cell Signaling Cat. 2772), BCL-2 (BD. Cat. 610539), BAK (Millipore Cat. 06-536), BIM (Cell Signaling Cat. 2933 S), Cleaved Caspase-3 (Cell Signaling Cat. 9664 S), Cleaved PARP (Cell Signaling Cat. 5625 S), COX-IV (Cell Signaling Cat. 4850 S), β-Actin (Sigma Cat. A1978), β-Tubulin (Cell Signaling Cat. 2146 S).
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