Sections of HCC samples and adjacent normal specimens were incubated with a rabbit polyclonal anti-GINS2 antibody (1:200, #ab197123, Abcam, Cambridge, UK). The results of IHC staining were examined under double-blinded conditions and scored on a semi-quantitative scale based on the score of intensity and extent. The score of intensity was marked as 0 (no staining), 1 (weak), 2 (moderate), and 3 (strong), and the percentage of positive stained area as 0 (none), 1 (1-25%), 2 (26-50%), 3 (51-75%), and 4 (>=75%). The IHC score was calculated by multiplying the score of intensity and extent in each sample (scale range 0-12). The levels of GINS2 were divided into “GINS2 Low” group (score <6) and “GINS2 High” group (scores >=6).
Ab197123
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab197123 is a laboratory equipment product from Abcam. It serves as a core function in scientific research and experiments, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ab181112, by Abcam (1 mentions)
Enhanced chemi luminescence (ecl), by Merck Group (1 mentions)
Ab6721, by Abcam (1 mentions)
Ripa assay buffer, by Beyotime (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Ecl reagent, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
3 protocols using ab197123
1
Immunohistochemical Analysis of GINS2 in HCC
2
Comparative Analysis of GINS Proteins in Pancreatic Cancer
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In this assessment, we aimed to compare the protein level of each GINS member between pancreatic cancer tissues and adjacent normal tissues. Also, the efficiency of gene transfection was detected. Proteins from pancreatic cancer tissues, adjacent normal tissues, and those correspondingly processed pancreatic cancer cells were extracted using the RIPA lysate. The protein concentration was determined via the BCA assay. Briefly, an equal amount of protein was loaded onto a sodium dodecyl sulfate (SDS) gel. Then, protein bands on the gel were transferred onto a polyvinylidene difluoride membrane, the membrane was blocked for 2 hours using 5% skim milk powder at room temperature. Thereafter, we incubated the membrane for 24 hours at 4°C with primary antibody for GINS1 (ab181112, 1:10,000, Abcam), GINS2 (ab197123, 1:1000, Abcam), GINS3 (ab177515, 1:2000, Abcam), GINS4 (ab139683, 1:1000, Abcam), respectively. Subsequently, the membrane was incubated for 1h at room temperature (RT) with secondary antibodies of the same species. Notably, GAPDH was used as the house-keeping gene. The blotting patterns were detected using ECL (Millipore) and visualized under an X-ray film.
3
Western Blot Analysis of GINS2 Protein
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After 48 h posttransfection, the cells were kept in a 6-well plate and managed under suitable conditions. After the cells were put into the RIPA assay buffer (Beyotime Biotech., Shanghai, China), protein in equal amounts was parted by SDS-PAGE and transferred onto PVDF membranes (Bio-Rad, CA, USA). Then, the membranes were blocked with skim milk (5%), cleansed with Tris-buffered saline, and incubated with primary antibodies, GINS2 (1 : 200, ab197123, Abcam, MA, USA) and GAPDH (1 : 1000, 5174T, Cell Signaling Technology, MA, USA), overnight at 4°C. After that, the membranes were incubated with secondary antibody HRP-conjugated goat antirabbit IgG heavy and light (1 : 2000, ab6721, Abcam) for 1 h. Furthermore, the membranes were analyzed by an ECL reagent (Thermo Scientific Pierce, IL, USA). The protein band was visualized with the internal reference of β-actin.
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