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G box imaging system

Manufactured by Merck Group
Sourced in United States

The G:BOX imaging system is a laboratory equipment designed for imaging and analysis of biological samples. It provides high-quality image capture and visualization capabilities for various applications, including gel electrophoresis, Western blots, and fluorescent imaging. The system is equipped with a sensitive camera and advanced optics to capture detailed images of samples, enabling researchers to document and analyze their experimental results.

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4 protocols using g box imaging system

1

Immunohistochemistry and Western Blot Analysis

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Immunohistochemistry (IHC) experiments and IHC scoring were carried out as previously described according to previous protocols [25 (link)]. Protein lysates were prepared by homogenization in 1% NP-40 containing 1 mM PMSF, and samples containing 20 μg of protein were separated by SDS-PAGE. The immunoreactive bands were visualized using an ImmobilonTM Western kit (Millipore, Billerica, MA) using the Syngene G:BOX imaging system (Frederick, USA). According to the manufacturer’s instructions, western blot assays were carried out with antibodies diluted in PBS plus Tween-20 (PBST). Information on all primary antibodies used is listed in Supplementary Table 2.
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2

Comprehensive Protein Expression Analysis

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The immunohistochemistry (IHC) and IHC scoring were carried out according to protocols that have been described previously [14 (link)]. Protein lysates were prepared by homogenization in 1% NP40 containing 1 mM PMSF and 20 μg protein was separated by SDS-PAGE. The immunoreactive bands were visualized using an Immobilon™ Western Kit (Millipore, Billerica, MA) using the SYNGENE G: BOX imaging system (Frederick, USA). Antibodies specific to CLDN7 (ab27487), BCL-2 (ab32124), PARP1 (ab32064), Caspase-3 (ab13847), E-cadherin (CDH1, ab76055), N-cadherin (CDH2, ab18203), Vimentin (ab92547) and TGFB1 (ab25121) were purchased from Abcam (Hong Kong, China). GAPDH (TA309157) and Ki-67 (TA500265) were purchased from ZSGB-BIO, Beijing, China. Cleaved-Caspase3 (Asp175) (9661S) was purchased from Cell Signaling Technology, MA, USA. The antibodies for the IHC and Western blot assays were diluted with phosphate buffered solution (PBS) or PBS plus Tween-20 (PBST), according to the manufacturer’s instructions.
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3

Protein Extraction and Western Blot

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Total protein was extracted from the cultured cells and specimens using RIPA buffer (ShineGene Molecular Biotech, Inc., Shanghai, China) according to the manufacturer's instructions. SDS-PAGE and Western blots were carried out following standard protocols. The immunoreactive bands were visualized by Immobilon™ Western Kit (Millipore, Billerica, MA) using a SYNGENE G: BOX imaging system (Frederick). The primary antibodies and the secondary HRP-conjugated antibodies were described in Table S2.
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4

CXCR4 Signaling in Hippocampal Tissue

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Hippocampal brain tissue was isolated using RIPA buffer containing PMSF (Bioteke Co., Ltd, China). On the basis of the molecular weight of the target protein, different concentrations of separation SDS-PAGE gels (Beyotime, China) and electrical conditions were prepared. Rabbit anti-CXCR4 and rabbit anti-p-PI3k antibodies (Abcam plc, UK) and rabbit anti-PI3k, rabbit anti-AKT and rabbit anti-p-AKT antibodies (Cell Signaling Technology, USA) were prepared. The proteins of interest were visualized using Luminata Crescendo Western HRP substrate (Millipore, Billerica, USA) and the Syngene G-box Imaging System.
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