Western blot analysis was used to evaluate the expression of AKT2, HK2, MMP2, STAT3, and p-STAT3 in AGS and SGC-7901 cells, respectively. A total of 5 primary antibodies including AKT2#2964, HK2#2867, MMP2#40994, STAT3#12640, and p-STAT3#9145 were purchased from Cell Signal Technology, Inc. (USA). GAPDH antibody was purchased from abMart Company (Shanghai, China). First, the band (expression intensity) was checked by Pierce™ ECL substrate then measured by ChemiDoc™ Touch Imaging System (Bio-Rad, USA). The GAPDH expression was detected in a similar sample as a control. The dilution of the primary antibody was performed based on the manufacturer’s instructions.
Pierce ecl substrate
Manufactured by Bio-Rad
Sourced in United States
The Pierce™ ECL substrate is a chemiluminescent detection reagent used for the sensitive detection of proteins in Western blot analysis. It utilizes a luminol-based reaction to produce a light signal proportional to the amount of target protein present in the sample.
Automatically generated - may contain errors
Lab products found in correlation
Stat3 12640, by Cell Signaling Technology (1 mentions)
P stat3 9145, by Cell Signaling Technology (1 mentions)
Mmp 2 40994, by Cell Signaling Technology (1 mentions)
Hk2 2867, by Cell Signaling Technology (1 mentions)
Chemidoc touch imaging system, by Bio-Rad (1 mentions)
Chemidoc touch gel imaging system, by Bio-Rad (1 mentions)
2 protocols using pierce ecl substrate
1
Protein Expression Analysis of Key Signaling Pathways
2
Protein Expression Analysis in HepG2 Cells
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HepG2 cells (5 × 105 cells/dish) were distributed in 100 mm culture dishes for 24 h. After 6 h of starvation, cells were incubated with cell growth medium containing R and Q for 24 h. Cell lysates were collected, and total protein was electrophoretically separated and transferred onto a PVDF membrane as described previously [49 (link)]. The membrane was incubated with primary antibody and HRP-conjugated relative secondary antibodies. Protein bands were visualized with Pierce ECL substrate and detected with the ChemiDoc Touch Gel Imaging System (Bio-Rad, Hercules, CA, USA); subsequently, the relative band intensity was quantified.
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