Lasmini 4000

Manufactured by GE Healthcare

Sourced in United Kingdom, United States

The LASmini 4000 is a compact and versatile laser system designed for precise material processing applications. It features a solid-state laser source that generates high-energy pulses, enabling efficient and accurate material removal, cutting, and welding. The system's small footprint and user-friendly interface make it suitable for a wide range of industrial and research settings.

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Lab products found in correlation

5 protocols using lasmini 4000

Protein Expression Analysis of Aortic Tissues

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We examined the protein expression of MMP-9, CD36, IL-6, and TNF-α in the upper abdominal aorta by Western blot analysis. In brief, 30 µg of tissue lysates were prepared as the standard protocol and separated on a 12% sodium dodecyl sulfate-polyacrylamide gel by electrophoresis. The protein bands were subsequently transferred to a polyvinylidene fluoride membrane for 1 hour and were blocked with 5% skimmed milk for 1 hour at room temperature. The membranes were then incubated with rat anti-MMP-9 (Abcam) and rat anti-CD36/IL-6/TNF-α (Bioss ANTIBODIES) at 4°C overnight. The membranes were subsequently incubated with human, anti-rabbit secondary antibodies (563-2; 1:1,000 dilution Zemai Biotech Corporation, Shanghai, China) for 1 hour at room temperature after three washes. Finally, the reaction was visualized using an enhanced chemiluminescence detection system (LAS MINI 4000, GE Healthcare Life Sciences, Little Chalfont, UK).

Li R., Cui S., Xu Y., Xing J., Xue L, & Chen Y. (2018). The upregulated scavenger receptor CD36 is associated with the progression of nontarget lesions after stent implantation in atherosclerotic rabbits. Journal of Inflammation Research, 11, 447-456.

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Western Blot Analysis of Apoptosis Markers

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Cells were harvested and lysed in SDS sample buffer (62.5 mM Tris-HCl, pH6.8, 10% glycerol, 2% SDS, 50 mM DTT and 0.01% bromphenol blue). Lysates were subjected to SDS-PAGE and transferred to PVDF membranes (Millipore). Membranes were blocked with 5% nonfat milk in Tris-buffered saline/0.1% Tween-20 and incubated at 4°C overnight with the following antibodies: anti-Axin 2, anti-Cyclin D1, anti-c-Myc, anti-Survivin, anti-caspase 3, anti-cleaved caspase 3, anti-caspase 8, anti-PARP-1 (Santa Cruz, CA), anti-Lamin A/C (Epitomics), anti-β-catenin (BD Biosciences), anti-phopho-β-catenin (Cell signaling Technology), and against β-actin (Santa Cruz, CA), followed by the corresponding horseradish peroxidase-conjugated secondary antibodies. Proteins of interest were visualized and imaged under chemi-luminescent detection using LASmini 4000 (GE Healthcare).

Kong L.M., Feng T., Wang Y.Y., Li X.Y., Ye Z.N., An T., Qing C., Luo X.D, & Li Y. (2016). Bisleuconothine A, a bisindole alkaloid, inhibits colorectal cancer cell in vitro and in vivo targeting Wnt signaling. Oncotarget, 7(9), 10203-10214.

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Whole Cell Lysate Protein Analysis

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For analysis of the whole cell lysate, SDS-PAGE buffer (62.5 mM Tris-HCl pH 6.8, 10% glycerol, 2% SDS, 50 mM DTT, and 0.01% bromophenol blue) was utilized to lyse cells after treatment. The subsequent SDS-PAGE and western blotting were carried out according to standard procedures [30 (link)]. Immunodetection was conducted using the following antibodies: anti-Survivin, anti-c-Myc, anti-Cyclin D1, anti-Axin 2, anti-PARP-1, anti-caspase 3, anti-cleaved caspase 3, anti-caspase 8 (Santa Cruz, CA, USA), anti-β-catenin (BD Biosciences), anti-phospho-β-catenin (Cell signaling Technology), anti-non-phospho (Active) β-catenin (Ser33/Ser37/Thr41) (Cell signaling Technology), anti-Lamin A/C (Epitomics), and anti-β-actin (Santa Cruz, CA, USA). Subsequently, the corresponding horseradish peroxidase (HRP)-conjugated secondary antibodies were used and protein bands were finally detected with LASmini 4000 (GE Healthcare, Barrington, IL, USA).

Ye Z.N., Yuan F., Liu J.Q., Peng X.R., An T., Li X., Kong L.M., Qiu M.H, & Li Y. (2019). Physalis peruviana-Derived 4β-Hydroxywithanolide E, a Novel Antagonist of Wnt Signaling, Inhibits Colorectal Cancer In Vitro and In Vivo. Molecules, 24(6), 1146.

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Germacrone-Induced Autophagy in Prostate Cancer

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Prostate cancer cells were treated with germacrone, 50 μM chloroquine (CQ) or 5 mM 3-methyladeseven (3-MA) was added to the culture medium for the last 6 h. After the cells were collected, the total proteins were extracted with RIPA buffer containing protease and phosphatase inhibitors on ice. The proteins were separated by SDS-PAGE and transferred to PVDF membranes. Then, the membrane was blocked with 5% BSA for 1 h. Subsequently, the PVDF membrane was incubated with primary antibodies overnight at 4°C and then incubated with an HRP-conjugated secondary antibody for 1 h. Protein bands were visualized using an enhanced chemiluminescence (ECL) kit. Images of the protein bands were captured by chemiluminescent detection (LASmini 4000, GE Healthcare).

Yu Z., Xu J., Shao M, & Zou J. (2020). Germacrone Induces Apoptosis as Well as Protective Autophagy in Human Prostate Cancer Cells. Cancer Management and Research, 12, 4009-4016.

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Western Blot Analysis of Apoptotic Markers

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Cells were harvested and lysed in SDS sample buffer (62.5 mM Tris-HCl, pH 6.8, 10% glycerol, 2% SDS, 50 mM DTT and 0.01% bromphenol blue). Lysates were subjected to SDS-PAGE and transferred to PVDF membranes (Millipore). Membranes were blocked with 5% nonfat milk in Tris-buffered saline/0.1% Tween-20 and incubated at 4 °C overnight with the following antibodies: anti-bcl-2 (Santa Cruz, CA), anti-bcl-xL (Santa Cruz, CA), anti-p65 (Santa Cruz, CA), anti-XIAP (Epitomics), anti-survivin (Santa Cruz, CA), anti-caspase 3 (Santa Cruz, CA), anti-cleaved caspase 3 (Santa Cruz, CA), anti-caspase 9 (Epitomics), anti-caspase 8 (Santa Cruz, CA), anti-PARP-1 (Santa Cruz, CA), anti-Lamin A/C (Epitomics) and against β-actin (Santa Cruz, CA), followed by the corresponding horseradish peroxidase-conjugated secondary antibodies. Proteins of interest were visualized and imaged under chemi-luminescent detection using LASmini 4000 (GE Healthcare).

Kong L.M., Deng X., Zuo Z.L., Sun H.D., Zhao Q.S, & Li Y. (2014). Identification and validation of p50 as the cellular target of eriocalyxin B. Oncotarget, 5(22), 11354-11364.

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