Genegnome hr image capture system

Manufactured by Syngene

Sourced in United States

The GeneGnome HR Image Capture System is a high-resolution imaging device designed for capturing images of biological samples, such as gels and blots. It provides a platform for capturing, analyzing, and documenting experimental results.

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Lab products found in correlation

Immobilon psq, by Merck Group (3 mentions) Complete mini, by Roche (2 mentions) Ecl supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Bicinchoninic acid protein assay kit, by Beyotime (1 mentions) Smad2 3, by Cell Signaling Technology (1 mentions) P smad2 3, by Cell Signaling Technology (1 mentions) P mek, by Cell Signaling Technology (1 mentions) P erk, by Cell Signaling Technology (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Anti rock1, by Abcam (1 mentions) Anti actin, by Abcam (1 mentions) Anti rhoa, by Abcam (1 mentions) Ripa buffer, by Beyotime (1 mentions) Bca protein assay kit, by Beyotime (1 mentions)

7 protocols using genegnome hr image capture system

Protein Expression Analysis in HUVECs

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Total proteins of HUVECs in 6-well plates were extracted using RIPA lysis buffer (Beyotime, Shanghai, China) supplemented with protease inhibitors (20 uL/mL) and phosphatase inhibitors (20 uL/mL). Then proteins (20ul samples per lane) were separated on 10% polyacrylamide gels by SDS-PAGE and transferred onto PVDF membranes. After blocked with 5% skimmed milk for 1 h.
At room temperature, the blots were incubated with various concentrations of primary antibodies at 4 °C overnight according to the manufacturer’s protocols. Then the membranes were washed three times in TBS-T for 5 min each time and subsequently incubated with horseradish peroxidase–conjugated goat anti-mouse or anti-rabbit antibodies (1:10,000 dilution) for 2 h, followed by three washes with TBS-T 5 to 7 min each time. Lastly, the blots incubated with ECL Super Signal West Pico Chemiluminescent Substrate (Thermo Scientific) and detected using the GeneGnome HR Image Capture System (Syngene).

Zhao N., Gui X., Fang Q., Zhang R., Zhu W., Zhang H., Li Q., Zhou Y., Zhao J., Cui X., Gao G., Tang H., Shen N., Chen T., Song H, & Shen W. (2022). Graphene quantum dots rescue angiogenic retinopathy via blocking STAT3/Periostin/ERK signaling. Journal of Nanobiotechnology, 20, 174.

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Protein Expression Analysis in Mouse Retina and HUVEC

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Lysis buffer containing protease inhibitor cocktail was used to extract total protein from mouse retinas and HUVECs. The protein concentration of each sample was determined using a bicinchoninic acid protein assay kit (Beyotime Institute of Biotechnology). Approximately 10 µg of protein was separated in 10% SDS/PAGE gels and transferred onto polyvinylidene fluoride membranes. Nonfat milk was used at 5% to block the non-specific sites. Subsequently, the samples were incubated with GAPDH, CD31, cyclin A1, cyclin B1, cyclin D1, cyclin E1, CDK1, CDK2, PARP1, p21 and p53 primary antibodies diluted at a ratio of 1:1,000 in Universal antibody diluent at 4°C overnight, following which the membranes were washed three times with TBS-T and incubated with HRP-conjugated secondary antibodies for 2 h at room temperature. GAPDH was used as a loading control and the protein signals were finally detected using Super Signal West Pico chemiluminescent substrate and semi-quantified using the GeneGnome HR Image Capture system (Syngene Europe).

Gao X., Zhang Y., Zhang R., Zhao Z., Zhang H., Wu J., Shen W, & Zhong M. (2019). Cyclin-dependent kinase 1 disruption inhibits angiogenesis by inducing cell cycle arrest and apoptosis. Experimental and Therapeutic Medicine, 18(4), 3062-3070.

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Western Blot Analysis of Signaling Proteins

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Protein extracts from HCC tissues or HepG2 cells were prepared using a modified RIPA buffer with 0.5% sodium dodecyl sulfate (SDS) in the presence of proteinase inhibitor cocktail (Complete mini, Roche, Indianapolis, IN, USA). Fifty micrograms of protein from HCC tissues and their adjacent non-tumor tissues were electrophoresed in 10% SDS-PAGE mini gels and transferred onto PVDF membranes (Immobilon P^-SQ, Millipore, Billerica, MA, USA). After blocking with 5% nonfat milk, the membranes were incubated with primary antibodies at 4°C overnight, followed by incubation with HRP-conjugated goat anti-rabbit or goat anti-mouse antibody (1:10000 dilution, KPL, Gaithersburg, MA,USA) for 1 hour at room temperature. Signals were developed with Super Signal West Pico chemiluminescent substrate (Pierce, Rockford, Il, USA), visualized using the Gene Gnome HR Image Capture System (Syngene, Frederick, MD, USA), and analyzed with Gene tools (Syngene). The primary antibodies were: MEK, p-MEK, ERK, p-ERK, SMAD2/3, p-SMAD2/3 (1:1000 dilution, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (1:5000 dilution, Epitomics Inc., Burlingame, CA, USA).

Xu G., Yang F., Ding C.L., Zhao L.J., Ren H., Zhao P., Wang W, & Qi Z.T. (2014). Small nucleolar RNA 113–1 suppresses tumorigenesis in hepatocellular carcinoma. Molecular Cancer, 13, 216.

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Western Blotting of Proteins from HUVECs

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Total protein of HUVECs was extracted in a modified Buffer with 0.5% SDS in the presence of proteinase inhibitor cocktail. Forty micrograms of proteins were electrophoresed in12.5% SDS/PAGE mini gels and transferred onto poly vinylidenedifluoride membranes. The nonspecific sites were blocked with 5% nonfat milk before the membranes were incubated with primary antibodies at 4℃ overnight. Then, the blots were washed 3 times for 5 to 7 minutes each time in TBS-T. Blots were incubated with horseradish peroxidaseconjugated antibodies for 2 hours at room temperature, followed by 3 washes of 5 to 7 minutes each time in TBS-T. Finally, blots were incubated with Super Signal West Pico chemoluminescent substrate and visualized using the GeneGnome HR Image Capture System (Syngene, Frederick, MD, USA).

Song H., Pan D., Sun W., Gu C., Zhang Y., Zhao P., Qi Z, & Zhao S. (2015). SiRNA directed against annexin II receptor inhibits angiogenesis via suppressing MMP2 and MMP9 expression. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 35(3).

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Western Blot Analysis of Fibrosis Markers

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Total protein was extracted using a modified buffer with 0.5% sodium dodecyl sulfate (SDS) containing a protease inhibitor cocktail (Sigma-Aldrich Co., St Louis, MO, USA). Approximately 20 µg of protein were electrophoresed using 10% SDS/polyacrylamide gel electrophoresis (PAGE) gels. The separated proteins were transferred to 0.22 µm poly-vinylidene fluoride membranes at 4°C. Nonfat milk (5%) was used to block nonspecific sites for 2 hours at room temperature. The membranes were then incubated with antibodies against GAPDH, α-SMA, TGF-β1, TGF-β2, Smad2, Smad3, p-Smad2 and p-Smad3 at 4°C overnight, followed by HRP-conjugated secondary antibodies for 2 hours. The membranes were washed three times in TBS-T, and bands were identified using enhanced chemiluminescence kits and visualized using the GeneGnome HR Image Capture System (Syngene, Frederick, MD, USA).

Zhao Z., Shen W., Zhu H., Lin L., Jiang G., Zhu Y., Song H, & Wu L. (2018). Zoledronate inhibits fibroblasts’ proliferation and activation via targeting TGF-β signaling pathway. Drug Design, Development and Therapy, 12, 3021-3031.

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Western Blot Analysis of MT1M in HCC

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Protein extracts from HCC tissues or HepG2 cells were prepared by a modified RIPA buffer with 0.5% sodium dodecyl sulfate (SDS) and proteinase inhibitor cocktail (Complete mini, Roche, Indianapolis, IN, USA). Fifty micrograms of protein were electrophoresed in 10% SDS-PAGE mini gels and transferred onto PVDF membranes (Immobilon P^−SQ, Millipore, Billerica, MA, USA). After blocking with 5% nonfat milk, the membranes were incubated with MT1M antibody (1:1000, Sigma-Aldrich, St. Louis, MO, USA) or GAPDH antibody (1:5000, Epitomics Inc., Burlingame, CA, USA at 4°C overnight, followed by incubation with HRP-conjugated goat anti-rabbit or goat anti-mouse antibody (1:10000 dilution, KPL, Gaithersburg, MA,USA) for 1 hour at room temperature. Finally, signals were developed with Super Signal West Pico chemoluminescent substrate (Pierce, Rockford, IL, USA), visualized by the Gene Gnome HR Image Capture System (Syngene, Frederick, MD, USA) and analyzed by Gene tools (Syngene).

Fu C.L., Pan B., Pan J.H, & Gan M.F. (2017). Metallothionein 1M suppresses tumorigenesis in hepatocellular carcinoma. Oncotarget, 8(20), 33037-33046.

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Western Blot Protein Analysis

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Cells were lysed using a RIPA buffer (Beyotime, Shanghai, China) with 0.5% sodium dodecyl sulfate in the presence of proteinase inhibitor cocktail (Complete mini; Roche). Then the protein concentration was measured using a BCA Protein Assay Kit (Beyotime). Twenty-five micrograms of protein were separated by 10% SDS-PAGE mini gels and then transferred onto a PVDF membrane (Immobilon P -SQ ;Millipore, Billerica, MA). After blocking with 5% nonfat milk, the membranes were incubated with primary anti-RhoA (1:1000, Abcam), anti-ROCK1 (1:1000,Abcam) or anti-actin (1:5000, Epitomics, Burlingame, CA) at 4°C overnight, followed by incubated with HRP-labeled secondary antibodies (anti-rabbit IgG, 1:10000, KPL, Gaithersburg, MA) for 1 h at room temperature. Finally, the band signals were developed with Super Signal West Pico chemoluminescent substrate (Pierce, Rockford, IL), visualized by the GeneGnome HR Image Capture System (Syngene, Frederick, MD) and analyzed by Gene tools (Syngene).

Yang L., Wang Z.F., Wu H, & Wang W. (2018). miR-142-5p Improves Neural Differentiation and Proliferation of Adipose-Derived Stem Cells. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 50(6).

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